| (11) | EP 3 234 135 B1 |
(12) | EUROPEAN PATENT SPECIFICATION |
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(54) | IMMUNOMODULATION BY CONTROLLING ELR+ PROINFLAMMATORY CHEMOKINE LEVELS WITH THE LONG NON-CODING RNA UMLILO IMMUNMODULATION DURCH KONTROLLE DER PROINFLAMMATORISCHEN ELR+CHEMOKINSPIEGEL MIT LONG-NON-CODING-RNA-UMLILO IMMUNOMODULATION PAR CONTRÔLE DES TAUX DE CHIMIOKINES PRO-INFLAMMATOIRES ELR+ AU MOYEN DU LONG ARN NON CODANT UMLILO |
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Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). |
BACKGROUND OF THE INVENTION
SUMMARY OF INVENTION
BRIEF DESCRIPTION OF THE FIGURES
Figure 1: UMLILO is a new super-enhancer resident IncRNA that is transcribed within the ELR+ CXC chemokine TAD: Hi-C analysis across the CXC chemokine locus reveals that the chemokine TAD spans a region of -500 Kbp.
Figure 2: UMLILO is a new super-enhancer resident lncRNA that is transcribed within the ELR+ CXC chemokine TAD: ChlA-PET data from a library constructed in HUVECs with an antibody that enriches for actively transcribing Pol II. Unstimulated HUVECs displayed relatively few PETs/chromosomal contacts. Upon chemokine gene induction with TNF for 30 mins numerous PETs/contacts were observed, occurring between the super-enhancer region and the chemokine genes, resulting in the formation of a putative 'inverted rosette' structure. Contacts detected by ChlA-PET between interacting chemokine genes, were confined exclusively to the region encompassing the chemokine TAD.
Figure 3: UMLILO is a new super-enhancer resident IncRNA that is transcribed within the ELR+ CXC chemokine TAD: ChlPseq analysis across the chemokine TAD revealed that the SE is atypically large (∼80 Kbp) and highly enriched for H3K4me1 and H3K27Ac epigenetic marks. The UMLILO IncRNA was identified from CAGE data to be expressed in unstimulated endothelial cells. UMLILO does not possess the typical eRNA histone modifications, H3K4me1 and H3K27Ac.
Figure 4: UMLILO transcription precedes chemokine gene activation: The chemokine TAD structure is formed prior to gene activation by TNF and UMLILO is brought in close proximity to the chemokine genes by pre-formed chromosomal loops.
Figure 5: UMLILO transcription precedes chemokine gene activation: Intronic smFISH reveals that the chemokines are only induced in HUVECs following TNF induction. Intronic RNA FISH foci of co-expressed chemokine genes always co-localize (n, number of alleles; scale bar, 5 µm; cells were counterstained with DAPI.).
Figure 6: UMLILO transcription precedes chemokine gene activation: RNA FISH on intron 1 and intron 2 of UMLILO reveal UMLILO is expressed in HUVECs prior to TNF stimulation. The signal from both introns consistently colocalized. Levels of intronic UMLILO were elevated shortly after TNF treatment, and then dropped back down to resting UMLILO levels after longer durations of TNF stimulation (n, number of alleles; scale bar, 5 µm; cells were counterstained with DAPI.).
Figure 7: UMLILO transcription precedes chemokine gene activation: Simultaneous intronic RNA FISH on UMLILO and the chemokine genes revealed that co-localization between intronic UMLILO and the chemokine genes was rarely observed (n, number of alleles; scale bar, 5 µm; cells were counterstained with DAPI.).
Figure 8: UMLILO transcription precedes chemokine gene activation: UMLILO is highly conserved in higher vertebrates with neutrophil rich blood, but no homolog of UMLILO or IL8 exists in mice.
Figure 9: UMLILO transcription is not influenced by perturbing transcriptional regulators that densely occupy the super-enhancer, but is necessary for chemokine transcription: The super-enhancer is densely occupied by chromatin regulators, including the mediator subunit, MED12, and the chromatin reader BRD4.
Figure 10: UMLILO transcription is not influenced by perturbing transcriptional regulators that densely occupy the super-enhancer, but is necessary for chemokine transcription: Chemokine expression is sensitive to BRD4 depletion. (+) JQ1 but not its enantiomer (-) JQ1 abrogated chemokine transcriptional activation, but did not influence UMLILO transcription (cells were counterstained with DAPI; scale bar, 5 µm).
Figure 11: UMLILO transcription is not influenced by perturbing transcriptional regulators that densely occupy the super-enhancer, but is necessary for chemokine transcription: Chemokine expression is sensitive to MED12 depletion. Depleting MED12 abrogates the expression of the CXC chemokines, but did not significantly alter the expression of UMLILO (Mean +/- SD; **p < 0.05, *** p < 0.01; two-tailed unpaired Student's T test).
Figure 12: UMLILO transcription is not influenced by perturbing transcriptional regulators that densely occupy the super-enhancer, but is necessary for chemokine transcription:UMLILO is necessary for chemokine expression. siRNA knockdown of UMLILO was sufficient to significantly abrogate chemokine expression, but not another TNF-induced gene located in a different TAD (Mean +/- SD; **p < 0.05, *** p < 0.01; two-tailed unpaired Student's T test).
Figure 13: UMLILO acts in cis to regulate chemokine expression: The CRISPR-Cas9 system was used to erase the genomic sequence encoding UMLILO and replace it with an eGFP reporter sequence in primary HUVEC cells.
Figure 14: UMLILO acts in cis to regulate chemokine expression: Successful repair was detected using RNA FISH probes targeting eGFP RNA, driven by the endogenous UMLILO promoter.
Figure 15: UMLILO acts in cis to regulate chemokine expression: Successful repair was observed in ∼26% of cells, with ∼19% of cells displaying a single foci, and ∼7% of cells displaying dual eGFP foci. Cells displaying a single eGFP focus were still able to express UMLILO from the opposite allele. The deletion of UMLILO abrogated chemokine expression in cis. Co-localization between eGFP foci and IL8 was never observed (Two-tailed Fisher's exact test; *p < 0.01; cells were counterstained with DAPI; Scale bar, 5 µm).
Figure 16: UMLILO acts in cis to regulate chemokine expression: UMLILO exerts its effect in cis. In cells displaying no eGFP foci, expression of IL8 and CXCL2 was comparable to the mock-transfected cells. In cells displaying a single eGFP focus, the expression of IL8 and CXCL2 was reduced significantly. Chemokine expression was never observed in cells displaying dual eGFP foci (Two-tailed Fisher's exact test; *p < 0.01; cells were counterstained with DAPI; Scale bar, 5 µm).
Figure 17: UMLILO interacts with the WAR complex to maintain H3K4me3 epigenetic regulation of chemokine genes: UMLILO interacts with WDR5. UMLILO was recovered from the nuclear fraction of cell lysates using biotinylated single stranded DNA probes tiling the exonic portion of the lncRNA. Using the MRMHR mass spectrophotometry approach, we identified WDR5, but not Ash2L, RbBP5 or DY30, in the UMLILO pull downs. Immunofluoresence revealed that WDR5 is a low abundant protein located in the nucleus. Immuno-RNA FISH revealed that distinct foci of WDR5 colocalize with UMLILO. Cyto.= cytoplasmic fraction (*p < 0.05, ** p < 0.01, two-tailed unpaired Student's T test; Scale bar, 5 µm).
Figure 18: UMLILO interacts with the WAR complex to maintain H3K4me3 epigenetic regulation of chemokine genes: UMLILO directs WDR5 to the chemokine promoters to facilitate their epigenetic activation. siUMLlLO modulates WDR5 binding as well as Pol II-Ser5 and H3K4me3 occupancy on chemokine promoters (*p < 0.05, ** p < 0.01, two-tailed unpaired Student's T test).
Figure 19: UMLILO interacts with the WAR complex to maintain H3K4me3 epigenetic regulation of chemokine genes: Targeting MLL1-WDR5 interactions abrogated chemokine transcription. MM102 led to a significant reduction in chemokine gene transcription (*p < 0.05, ** p < 0.01, two-tailed unpaired Student's T test).
Figure 20: UMLILO does not regulate chromosomal contact across the chemokine TAD: Depleting UMLILO using siRNA does not significantly alter chromosomal contact across the chemokine TAD. ChlA-PET data reveals the sites of active Pol II-mediated chromosomal contacts.
Figure 21: UMLILO does not regulate chromosomal contact across the chemokine TAD: siUMLlLO did not significantly alter the chromosomal contacts between UMLILO and IL8. siGFP was used as a control for the detection of baseline chromosomal contacts.
Figure 22: The activity of UMLILO can be substituted with a different WDR5-interacting enhancer-like lncRNA: HOTTIP FISH spots were observed at approximately 25% of alleles and only single allelic expression of HOTTIP per HeLa cell was observed (n, number of cells; cells were counterstained with DAPI; Scale bar, 5 µm).
Figure 23: The activity of UMLILO can be substituted with a different WDR5-interacting enhancer-like lncRNA: Graphical representation of the repair strategy. The repair construct consists of the sequence of HOTTIP flanked by homologous arms that flank UMLILO.
Figure 24: The activity of UMLILO can be substituted with a different WDR5-interacting enhancer-like lncRNA: Replacing endogenous UMLILO with HOTTIP restores chemokine transcription. Overlapping HOTTIP and chemokine foci were observed in ∼1% of HeLas dual transfected with the CRISPR and HOTTIP repair construct for 120 hr and stimulated with TNF for 24 hrs (n, number of cells; cells were counterstained with DAPI; Scale bar, 5 µm).
Figure 25: The activity of UMLILO can be substituted with a different WDR5-interacting enhancer-like IncRNA: WDR5-interacting IncRNAs exploit pre-formed 3D chromatin folding to coordinate rapid chemokine gene activation. By acting in cis, WDR5-interacting lncRNAs use the local chromatin compaction of the pre-formed chemokine TAD to direct the WDR5 protein across the chemokine promoters. Upon activation by TNF, the chemokine super-enhancer becomes loaded with BRD4 and other transcriptional regulators, to induce the rapid and robust transcriptional induction of the chemokine genes.
Figure 26: UMLILO is a novel lncRNA of the human ELR+ chemokine locus that binds directly to the WDR5 protein to recruit the MLL H3K4 methylase complex, maintaining H3K4me3 on the chemokine promoters.
Figure 27: Tacrolimus inhibits UMLILO and IL-8 expression. The LASAGNA (Length-Aware Site Alignment Guided by Nucleotide Association) algorithm for transcription factor binding site (TFBS) alignment (http://biogrid.engr.uconn.edu/ lasagna_search/) was used to analyze the UMLILO promoter for putative TFBS. A high confidence binding site for the calcium dependent transcription factors, NF-AT1 and NF-AT2 was observed near to the TSS of UMLILO.
Figure 28: Tacrolimus inhibits UMLILO and IL-8 expression. CHIP assays confirm that the NF-AT transcription factors occupy the UMLILO promoter.
Figure 29: Tacrolimus inhibits UMLILO and IL-8 expression. Tacroliumus, a macrolide calcium inhibitor, leads to a reduction in UMLILO and IL-8 expression.
DETAILED DESCRIPTION OF THE INVENTION
Statistics
EXAMPLE 1
Cell Culture
Hi-C analysis
ChlA-PET analysis
Identification of enhancers in the chemokine TAD
Identification of long noncoding RNAs in the chemokine TAD
CRISPR synthesis
RNA interference
UMLILO siRNA 1: 5' AUC UUA AAU UAG AGG CGA AUU 3' (SEQ ID NO:54)
UMLILO siRNA 2: 5' CAU ACA AAU UCU CGC AGC AUU 3' (SEQ ID NO:55)
UMLILO siRNA 3: 5' AAG AGU UGG UAC ACG GUG AUU 3' (SEQ ID NO:56)
UMLILO siRNA 4: 5' GCA UAU UAA CCC UAC AAG UUU 3' (SEQ ID NO:57)
RT-qPCR
Target | Direction | Primer sequence | SEQ ID NO. |
UMLILO intron 1 | FW | 5' GGAAGGAGGGGAACATGGAA 3' | SEQ ID NO:2 |
RW | 5' CTACCTGACTCCCTCCCTCT 3' | SEQ ID NO:3 | |
UMLILO exon 3 | FW | 5' GGTACACGGTGAACATTTGCT 3' | SEQ ID NO:4 |
RW | 5' CAGCATCTCTCTGTCCACTGA 3' | SEQ ID NO:5 | |
IL8 gene | FW | 5' AGGGCCAAGAGAATATCCGA 3' | SEQ ID NO:6 |
RW | 5' GGACTTGTGGATCCTGGCTA 3' | SEQ ID NO:7 | |
CXCL1 gene | FW | 5' AGCTTGCCTCAATCCTGCAT 3' | SEQ ID NO:8 |
RW | 5' CCTCCTCCCTTCTGGTCAGT 3' | SEQ ID NO:9 | |
CXCL2 gene | FW | 5' CACAGTGTGTGGTCAACATTTCTC 3' | SEQ ID NO:10 |
RW | 5' ACACAGAGGGAAACACTGCATAA 3' | SEQ ID NO:11 |
RNA FISH probes
Immuno-RNA FISH
Image acquisition and processing
Repair construct
FW: 5' TTGAACCGGGTTTTCCAGTCACATATGGTGAGCAAGGGCGA 3' (SEQ ID NO:12)
RW: 5' ACTATGAAGACTCTTGGGTCACTTGTACAGCTCGTCCA 3' (SEQ ID NO:13)
Chromatin immunoprecipitation
Target | Di rection | Primer sequence | SEQ ID NO. |
IL8 | FW | 5' TGGGCCATCAGTTGCAAATC 3' | SEQ ID NO:14 |
RW | 5' AGTGAGATGGTTCCTTCCGG 3' | SEQ ID NO:15 | |
CXCL1 | FW | 5' ACGTGGGTCTAAGGGATCTG 3' | SEQ ID NO:16 |
RW | 5' GGGTCTGACTGTCTTGCGTA 3' | SEQ ID NO:17 | |
CXCL2 | FW | 5' CTGTGGTGGTTCTCAGGGAT 3' | SEQ ID NO:18 |
RW | 5' TGGACTCTGAGACTCTGGGA 3' | SEQ ID NO:19 | |
CXCL3 | FW | 5' TCTGGAATCCGAGACGATGG 3' | SEQ ID NO:20 |
RW | 5' GACAGGAAAGGCACGACTTC 3' | SEQ ID NO:21 | |
SAMD4A | FW | 5' GAGCTTTGGGTGGAGAGAGT 3' | SEQ ID NO:22 |
RW | 5' TTACTCCTCCTCCTCCTCCC 3' | SEQ ID NO:23 | |
UMLILO | FW | 5' TGTCCAAATCCACATTGACAGT 3' | SEQ ID NO:24 |
RW | 5' GGAGTGTTGCTGCGAGAATT 3' | SEQ ID NO:25 |
RNA immunoprecipitation with targeted mass spectrophotometry
3C assay
Primer | Fragment location | Sequence | SEQ ID NO |
Anchor | chr4:74604292-74606315 | 5' TCCTCTGACATAATGAAAAGATGAGGGTGC 3' | SEQ ID NO:26 |
1 | chr4:74499669-74501601 | 5' AAAATAGAAACCCTGAATGTACCGGTAACA 3' | SEQ ID NO:27 |
2 | chr4:74511769-74524359 | 5' TCAAATCCGTGATCAGCATTACCAAGCCAT 3' | SEQ ID NO:28 |
3 | chr4:74527725-74529421 | 5' ATTGGAAGGCTAAATATACTTACATGGC 3' | SEQ ID NO:29 |
4 | chr4:74530103-74534800 | 5' TTTTAAAAGCAGCACTAGTGTATCCGG 3' | SEO ID NO:30 |
5 | chr4:74546061-74548099 | 5' AAAGAGCTCAATGTGTGTTACTAAAGAATG 3' | SEQ ID NO:31 |
6 | chr4:74549704-74551014 | 5' AAAGGTAAGCAATAATTGGCCCATATCTC 3' | SEQ ID NO:32 |
7 | chr4:74553813-74555419 | 5' AAAAGTGATACATGTTCATTGCATTTAAAC 3' | SEQ ID NO:33 |
8 | chr4:74578660-74585594 | 5' CGTAAGGCTGGTCAAGGTATGCTGAG 3' | SEQ ID NO:34 |
9 | chr4:74585594-74587345 | 5' TTCTTCATTATGCTAGTGTTGTGTATTGTG 3' | SEQ ID NO:35 |
10 | chr474587345-74591147 | 5' TGAAATTAGAGAAAAACATGTACTTAGGGA 3' | SEQ ID NO:36 |
11 | chr4:74591147-74596339 | 5' CACCCCGCCCATTTGATGAACTGTTT 3' | SEQ ID NO:37 |
12 | chr4:74596459-74601876 | 5' ACTTCAAACTCCAAACTCCACCGATTTG 3' | SEQ ID NO:38 |
13 | chr4:74601963-74604292 | 5' CCAACATCACTGAAGCAAAGAAACTTGGAG 3' | SEQ ID NO:39 |
14 | chr4:74607958-74612686 | 5' TCTGCTTGTACGTAGGTATGTAGATT 3' | SEQ ID NO:40 |
15 | chr4:74722664-74725426 | 5' GCTAGCAACCAACTCTTTAAGAATACAGCC 3' | SEQ ID NO:41 |
16 | chr4:74736063-74750395 | 5' AGAAACACAACGATACAATGTGAAA 3' | SEQ ID NO:42 |
17 | chr4:74751595-74756828 | 5'AGTGTGACTCCAAATAACCTAGTTTGCTA 3' | SEQ ID NO:43 |
18 | chr4:74756828-74759327 | 5' ACACAGTCATAATCACAACCCCAGTC 3' | SEQ ID NO:44 |
19 | chr4:74899318-74900128 | 5' TCAGACTCATGGGCTCAGTTGATTC 3' | SEQ ID NO:45 |
20 | chr4:74901502-74902795 | 5' GGCTGACACATTATGGTCTCCCACTAAATA 3' | SEQ ID NO:46 |
21 | chr4:74903866-74908292 | 5' GAAACATGTCAAGAGGCCGTGGACATTT 3' | SEQ ID NO:47 |
22 | 74917831-74918715 | 5' AGTGTGAAACAAAACGAGAAGGGAAG 3' | SEQ ID NO:48 |
23 | 74964374-74967509 | 5' AAATTATTTGCTTTAGGAAGGGAAGTAGAA 3' | SEQ ID NO:49 |
24 | 74972881-74981053 | 5' CATAGGAGAGACTGCGACAGAAATTCCATT 3' | SEQ ID NO:50 |
25 | 74981053-74983881 | 5' TTACAACTCCTACAACCGTGCTTGGTACAT 3' | SEQ ID NO:51 |
GAPDH control F1 | chr12:6639882-6641136 | 5' TGCCAATCTCCTTGTTTTCTAATG 3' | SEQ ID NO:52 |
GAPDH control F2 | chr12:6641136-6645917 | 5' TATTCCCCCAGGTTTACATGTTC 3' | SEQ ID NO:53 |
EXAMPLE 2
The ELR+ CXC chemokines engage in pre-formed chromosomal contact
UMLILO is a super-enhancer resident IncRNA transcribed within the chemokine TAD
EXAMPLE 3
UMLILO transcription precedes chemokine gene activation
UMLILO is conserved in higher vertebrates, but lacks a homolog in rodents
EXAMPLE 4
UMLILO transcription is impervious to perturbation of super-enhancer transcriptional regulators and is necessary for chemokine transcription
EXAMPLE 5
UMLILO acts in cis to regulate chemokine expression
EXAMPLE 6
UMLILO interacts with the WAR complex to maintain H3K4me3 epigenetic regulation of chemokine genes
EXAMPLE 7
Depleting UMLILO does not abrogate chromosomal contact across the chemokine TAD
EXAMPLE 8
The activity of UMLILO can be substituted with a different WDR5-interacting IncRNA
Discussion
REFERENCES
Cong, L, et al. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819-23.
SEQUENCE LISTING
<110> CSIR
<120> IMMUNOMODULATION BY CONTROLLING ELR+ PROINFLAMMATORY CHEMOKINE LEVELS WITH THE LONG NON-CODING RNA UMLILO
<130> PA162147/PCT
<150> ZA 2014/09351
<151> 2014-12-18
<160> 314
<170> PatentIn version 3.5
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ggtacacggt gaacatttgc t 21
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cagcatctct ctgtccactg a 21
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agggccaaga gaatatccga 20
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ggacttgtgg atcctggcta 20
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agcttgcctc aatcctgcat 20
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cctcctccct tctggtcagt 20
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cacagtgtgt ggtcaacatt tctc 24
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acacagaggg aaacactgca taa 23
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ttgaaccggg ttttccagtc acatatggtg agcaagggcg a 41
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actatgaaga ctcttgggtc acttgtacag ctcgtcca 38
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<223> ChIP IL8 forward primer
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tgggccatca gttgcaaatc 20
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<223> ChIP IL8 reverse primer
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agtgagatgg ttccttccgg 20
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ctgtggtggt tctcagggat 20
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tggactctga gactctggga 20
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tctggaatcc gagacgatgg 20
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gacaggaaag gcacgacttc 20
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gagctttggg tggagagagt 20
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ttactcctcc tcctcctccc 20
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<400> 24
tgtccaaatc cacattgaca gt 22
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<223> ChIP UMLILO reverse primer
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ggagtgttgc tgcgagaatt 20
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<223> 3C anchor
<400> 26
tcctctgaca taatgaaaag atgagggtgc 30
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<223> 3C primer 1
<400> 27
aaaatagaaa ccctgaatgt accggtaaca 30
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<400> 28
tcaaatccgt gatcagcatt accaagccat 30
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attggaaggc taaatatact tacatggc 28
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ttttaaaagc agcactagtg tatccgg 27
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aaagagctca atgtgtgtta ctaaagaatg 30
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aaaggtaagc aataattggc ccatatctc 29
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aaaagtgata catgttcatt gcatttaaac 30
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cgtaaggctg gtcaaggtat gctgag 26
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ttcttcatta tgctagtgtt gtgtattgtg 30
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tgaaattaga gaaaaacatg tacttaggga 30
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caccccgccc atttgatgaa ctgttt 26
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acttcaaact ccaaactcca ccgatttg 28
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ccaacatcac tgaagcaaag aaacttggag 30
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tctgcttgta cgtaggtatg tagatt 26
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gctagcaacc aactctttaa gaatacagcc 30
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agaaacacaa cgatacaatg tgaaa 25
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tcagactcat gggctcagtt gattc 25
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ggctgacaca ttatggtctc ccactaaata 30
<210> 47
<211> 28
<212> DNA
<213> Artificial sequence
<220>
<223> 3C primer 21
<400> 47
gaaacatgtc aagaggccgt ggacattt 28
<210> 48
<211> 26
<212> DNA
<213> Artificial sequence
<220>
<223> 3C primer 22
<400> 48
agtgtgaaac aaaacgagaa gggaag 26
<210> 49
<211> 30
<212> DNA
<213> Artificial sequence
<220>
<223> 3C primer 23
<400> 49
aaattatttg ctttaggaag ggaagtagaa 30
<210> 50
<211> 30
<212> DNA
<213> Artificial sequence
<220>
<223> 3C primer 24
<400> 50
cataggagag actgcgacag aaattccatt 30
<210> 51
<211> 30
<212> DNA
<213> Artificial sequence
<220>
<223> 3C primer 25
<400> 51
ttacaactcc tacaaccgtg cttggtacat 30
<210> 52
<211> 24
<212> DNA
<213> Artificial sequence
<220>
<223> 3C GAPDH control F1
<400> 52
tgccaatctc cttgttttct aatg 24
<210> 53
<211> 23
<212> DNA
<213> Artificial sequence
<220>
<223> 3C GAPDH control F2
<400> 53
tattccccca ggtttacatg ttc 23
<210> 54
<211> 21
<212> RNA
<213> Artificial sequence
<220>
<223> UMLILO siRNA 1
<400> 54
aucuuaaauu agaggcgaau u 21
<210> 55
<211> 21
<212> RNA
<213> Artificial sequence
<220>
<223> UMLILO siRNA 2
<400> 55
cauacaaauu cucgcagcau u 21
<210> 56
<211> 21
<212> RNA
<213> Artificial sequence
<220>
<223> UMLILO siRNA 3
<400> 56
aagaguuggu acacggugau u 21
<210> 57
<211> 21
<212> RNA
<213> Artificial sequence
<220>
<223> UMLILO siRNA 4
<400> 57
gcauauuaac ccuacaaguu u 21
<210> 58
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 1
<400> 58
ttgtcctaga agcttgtgtg 20
<210> 59
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 2
<400> 59
cacagtgaga tggttccttc 20
<210> 60
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 3
<400> 60
cagcttggaa gtcatgttta 20
<210> 61
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 4
<400> 61
gaaatcagga aggctgccaa 20
<210> 62
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 5
<400> 62
cttaccttca cacagagctg 20
<210> 63
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 6
<400> 63
acgctgtagg tcagaaagat 20
<210> 64
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 7
<400> 64
ggatcacatt tagacatagg 20
<210> 65
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 8
<400> 65
caagaatagc tttgctatct 20
<210> 66
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 9
<400> 66
aggatgtttg ttaccaaagc 20
<210> 67
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 10
<400> 67
ctttatactt tcctaagtgt 20
<210> 68
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 11
<400> 68
atatatgcat gctacatggt 20
<210> 69
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 12
<400> 69
tataggtaac cgtggttctc 20
<210> 70
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 13
<400> 70
ccttgctcaa ctcaatatta 20
<210> 71
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 14
<400> 71
aggagtggaa ttgtctgagt 20
<210> 72
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 15
<400> 72
gacaaatctg aggcttgtca 20
<210> 73
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 16
<400> 73
gggtatcttt aaaccagaca 20
<210> 74
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 17
<400> 74
aagttataca tgcctggtct 20
<210> 75
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 18
<400> 75
gcatagaaat tgccttcaga 20
<210> 76
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 19
<400> 76
gtaggcaagc taagactctc 20
<210> 77
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 20
<400> 77
gtgataccac agtgttattt 20
<210> 78
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 21
<400> 78
ggtcaatatt gcataatcct 20
<210> 79
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 22
<400> 79
tcaacatctt catggtattt 20
<210> 80
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 23
<400> 80
gttagagttc tttttgtaca 20
<210> 81
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 24
<400> 81
acaacattga acgacttcct 20
<210> 82
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 25
<400> 82
cctcagttag ttctttgttt 20
<210> 83
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 26
<400> 83
tgaatattct cctagccctt 20
<210> 84
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 27
<400> 84
catgtttcta gtgaactcat 20
<210> 85
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 28
<400> 85
atttggagac tatggaaggc 20
<210> 86
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 29
<400> 86
ctacttcctt tctaattcca 20
<210> 87
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 30
<400> 87
tttatcaaca ggcacagctc 20
<210> 88
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 31
<400> 88
cacctgttgg gggaaaaagt 20
<210> 89
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 32
<400> 89
ttagcactcc ttggcaaaac 20
<210> 90
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 33
<400> 90
gtctttatgc actgacatct 20
<210> 91
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 34
<400> 91
tttggggtgg aaaggtttgg 20
<210> 92
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 35
<400> 92
tctcaatcac tctcagttct 20
<210> 93
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 36
<400> 93
cataatttct gtgttggcgc 20
<210> 94
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 37
<400> 94
ttccaccttc cttaatttta 20
<210> 95
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 95
<400> 95
cacctggaac tttcctaaat 20
<210> 96
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 39
<400> 96
tcatttacta ctgtaatcct 20
<210> 97
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 40
<400> 97
tagtagttgt ggaagctacc 20
<210> 98
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 41
<400> 98
aggcataaag tctaagtcca 20
<210> 99
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 42
<400> 99
gcattcaaat catgattcct 20
<210> 100
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 43
<400> 100
tctcttccat cagaaagctt 20
<210> 101
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> IL-8 smFISH probe 44
<400> 101
aaaacttctc cacaaccctc 20
<210> 102
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 1
<400> 102
aagcagccag gagcaaactc 20
<210> 103
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 2
<400> 103
tggagtagca gcactgacat 20
<210> 104
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 3
<400> 104
ggaaagagcg acacttacct 20
<210> 105
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 4
<400> 105
atagtctttc ttctgtgcta 20
<210> 106
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 5
<400> 106
cagttgagct caaggctaaa 20
<210> 107
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 6
<400> 107
taacagtttt ccatccctta 20
<210> 108
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 7
<400> 108
tcactacctc ttgtatactt 20
<210> 109
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 8
<400> 109
caccatgggc agacaacttc 20
<210> 110
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 9
<400> 110
cagcagtaac tcttctctcc 20
<210> 111
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 10
<400> 111
gcagagagga aagcctgatc 20
<210> 112
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 11
<400> 112
cccaaatctg ctgagattga 20
<210> 113
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 12
<400> 113
tagggatctc taagagccca 20
<210> 114
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 13
<400> 114
actttggctt tatcctgaag 20
<210> 115
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 14
<400> 115
ttatatcact cccagaaaca 20
<210> 116
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 15
<400> 116
tctactggta acattacaca 20
<210> 117
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 16
<400> 117
tctacatttc aaaatggccc 20
<210> 118
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 17
<400> 118
agttggggac acaggaggat 20
<210> 119
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 18
<400> 119
ctattaagtt gggaggtacg 20
<210> 120
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 19
<400> 120
gggatgttga tgtcaggtat 20
<210> 121
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 20
<400> 121
aaagttggca agcaatgtcc 20
<210> 122
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 21
<400> 122
acatgctgta aggtaaggga 20
<210> 123
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 22
<400> 123
aatacttgga cattcccgag 20
<210> 124
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 23
<400> 124
gtgttgtgtg actattgagc 20
<210> 125
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 24
<400> 125
gcatttttag gaatgcacct 20
<210> 126
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 25
<400> 126
ctggctacta agtattgaga 20
<210> 127
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 26
<400> 127
tgcttgaagg aacccttatg 20
<210> 128
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 27
<400> 128
ttcaagaata tgagcagccc 20
<210> 129
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 28
<400> 129
gtgggatatc ttcagtaaga 20
<210> 130
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 29
<400> 130
gagaaacaca ggacttctca 20
<210> 131
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 30
<400> 131
aacagtttgt gtgccgagaa 20
<210> 132
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 31
<400> 132
ggtcgttgag aaccaatcta 20
<210> 133
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 32
<400> 133
aatgtcttgg aaacctttgt 20
<210> 134
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 33
<400> 134
aaccacttag aggaggagtt 20
<210> 135
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 34
<400> 135
acttatcctg tgattggtat 20
<210> 136
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 35
<400> 136
aggtattgga tccacatgaa 20
<210> 137
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 36
<400> 137
gacagcagtc aaagttgctt 20
<210> 138
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 37
<400> 138
agaatacggt ctgtgtatcc 20
<210> 139
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 38
<400> 139
cgtgtgaagc ccacaataaa 20
<210> 140
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 39
<400> 140
gatagcattg atgtcacagc 20
<210> 141
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 40
<400> 141
agtttctagg tattcctgat 20
<210> 142
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 41
<400> 142
tttgccacat catttctttc 20
<210> 143
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 42
<400> 143
aaaggttttg ccttattcca 20
<210> 144
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 43
<400> 144
actagttccg tttgaccaaa 20
<210> 145
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 44
<400> 145
tagaaagtga tttttgccct 20
<210> 146
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 45
<400> 146
atcttcccta ttttcctaat 20
<210> 147
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 46
<400> 147
ctttctggta gaagatgctc 20
<210> 148
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CCL20 smFISH probe 47
<400> 148
agattttact tttggtctca 20
<210> 149
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 1
<400> 149
agggaatctc gtgagacagg 20
<210> 150
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 2
<400> 150
cattcttgag tgtggctctg 20
<210> 151
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 3
<400> 151
gggttgagac aagctttctg 20
<210> 152
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 4
<400> 152
tttcttaacc atgggcgatg 20
<210> 153
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 5
<400> 153
ctttttcagc atcttttcga 20
<210> 154
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 6
<400> 154
cgcctgtgta catggaaatt 20
<210> 155
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 7
<400> 155
gtatttctga ccaacagctc 20
<210> 156
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 8
<400> 156
actttttagg gggcagacgc 20
<210> 157
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 9
<400> 157
taaccctggg ttttcctgat 20
<210> 158
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 10
<400> 158
ataatttttt cagtcctgca 20
<210> 159
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 11
<400> 159
ccttaatggc aactttgtga 20
<210> 160
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 12
<400> 160
tctggcacca gaacagatta 20
<210> 161
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 13
<400> 161
aggagggaag aacctgggtg 20
<210> 162
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 14
<400> 162
gcagttttac ctacattcac 20
<210> 163
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 15
<400> 163
tccaatgaga ccttacatga 20
<210> 164
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 16
<400> 164
cacttcagca caactggcaa 20
<210> 165
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 17
<400> 165
gtgtttttta acaggtggaa 20
<210> 166
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 18
<400> 166
ggaactaggg gaaatgaagc 20
<210> 167
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 19
<400> 167
gcttttcagg tctgaaagtc 20
<210> 168
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 20
<400> 168
tttacaaaaa accagggggt 20
<210> 169
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 21
<400> 169
attgctcttt gtcatactgg 20
<210> 170
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 22
<400> 170
ctggctaaaa cagccagagc 20
<210> 171
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 23
<400> 171
agtggcagtc tacactgttt 20
<210> 172
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 24
<400> 172
tcttttagga atgggggtga 20
<210> 173
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 25
<400> 173
tgtgtggcaa ggacctctag 20
<210> 174
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 26
<400> 174
aagtgggtac tgcctgtgac 20
<210> 175
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 27
<400> 175
gttttctgaa tgcagtttcc 20
<210> 176
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 28
<400> 176
cacaacagtc ttctcctgtc 20
<210> 177
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 29
<400> 177
taggtgctca ggaaagctga 20
<210> 178
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 30
<400> 178
aatgagaaaa tgggtgccct 20
<210> 179
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 31
<400> 179
cttctggtca gttggatttg 20
<210> 180
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 32
<400> 180
aacagccacc aataagcttc 20
<210> 181
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 33
<400> 181
ctcttctgtt cctgtaaggg 20
<210> 182
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 34
<400> 182
aacacattag gcgcaatcca 20
<210> 183
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 35
<400> 183
agacttctcc taagtgatgc 20
<210> 184
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 36
<400> 184
gggagagtgt gcaagtagat 20
<210> 185
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 37
<400> 185
actaacttgg gtttgaccta 20
<210> 186
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL2 smFISH probe 38
<400> 186
ctgcaaacct tctatcttca 20
<210> 187
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 1
<400> 187
tcactgactg agcgaggctg 20
<210> 188
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 2
<400> 188
acgctcctag ggaagaagag 20
<210> 189
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 3
<400> 189
atgttcttgg ggtgaattcc 20
<210> 190
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 4
<400> 190
ggacttcacg ttcacacttt 20
<210> 191
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 5
<400> 191
ggacttacat gacttcggtt 20
<210> 192
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 6
<400> 192
agggaaggga atctcgtgag 20
<210> 193
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 7
<400> 193
cattcttgag tgtggctctg 20
<210> 194
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 8
<400> 194
caggattgag gcaagctttc 20
<210> 195
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 9
<400> 195
attttcttaa ctatggggga 20
<210> 196
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 10
<400> 196
taactcacct gttcagcatc 20
<210> 197
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 11
<400> 197
tcgcctgtgt acatggaaac 20
<210> 198
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 12
<400> 198
cagtatttct gaccaacggc 20
<210> 199
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 13
<400> 199
ttttattttt agggggcaca 20
<210> 200
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 14
<400> 200
caactaaccc ctgggttttc 20
<210> 201
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 15
<400> 201
ataatcccaa tttctagtcc 20
<210> 202
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 16
<400> 202
gcaccagagc agtaattatc 20
<210> 203
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 17
<400> 203
caggcattgg gaatatcctc 20
<210> 204
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 18
<400> 204
cgaaggaaga accagggtgg 20
<210> 205
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 19
<400> 205
gttttaccta cattcattgg 20
<210> 206
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 20
<400> 206
tactgggcct caaatgaagg 20
<210> 207
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 21
<400> 207
aaacagccag aacctgttgc 20
<210> 208
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 22
<400> 208
tctccactgt actattggat 20
<210> 209
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 23
<400> 209
cttttaggaa tgggggtggg 20
<210> 210
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 24
<400> 210
ggacctctaa gcttgggatg 20
<210> 211
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 25
<400> 211
tatgacagct gtgctgtgtg 20
<210> 212
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 26
<400> 212
tggcaaccaa gtggctactg 20
<210> 213
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 27
<400> 213
agagttttcc gaatgcagtt 20
<210> 214
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 28
<400> 214
attgcacaac actcttctcc 20
<210> 215
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 29
<400> 215
gagtaggtgc tcgggaaagc 20
<210> 216
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 30
<400> 216
gcaatgagaa aatgggtgcc 20
<210> 217
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 31
<400> 217
cttctggtca gttggatttg 20
<210> 218
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 32
<400> 218
ttcaggaaca gccaccagtg 20
<210> 219
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 33
<400> 219
ctcttctgtt cctataaggg 20
<210> 220
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 34
<400> 220
acacattagg cacaatccag 20
<210> 221
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 35
<400> 221
agaagacttc tcctaagcga 20
<210> 222
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 36
<400> 222
cccttaatta ttttacacct 20
<210> 223
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 37
<400> 223
ataggacagt gtgcaggtag 20
<210> 224
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 38
<400> 224
acttggggtt gacatttcaa 20
<210> 225
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 39
<400> 225
gcaggctcct cagaaatatt 20
<210> 226
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 40
<400> 226
ctatcacagt ggctggcatg 20
<210> 227
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> CXCL1 smFISH probe 41
<400> 227
atgatctcat tggccatttg 20
<210> 228
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 1
<400> 228
tcctcgccct tgctcaccat 20
<210> 229
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 2
<400> 229
atgggcacca ccccggtgaa 20
<210> 230
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 3
<400> 230
ttacgtcgcc gtccagctcg 20
<210> 231
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 4
<400> 231
gacacgctga acttgtggcc 20
<210> 232
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 5
<400> 232
ggcatcgccc tcgccctcgc 20
<210> 233
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 6
<400> 233
tcagggtcag cttgccgtag 20
<210> 234
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 7
<400> 234
ttgccggtgg tgcagatgaa 20
<210> 235
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 8
<400> 235
ggtgggccag ggcacgggca 20
<210> 236
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 9
<400> 236
cgtaggtgaa ggtggtcacg 20
<210> 237
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 10
<400> 237
tagcggctga agcactgcac 20
<210> 238
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 11
<400> 238
gtgctgcttc atgtggtcgg 20
<210> 239
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 12
<400> 239
gcatggcgga cttgaagaag 20
<210> 240
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 13
<400> 240
cgctcctgga cgtagccttc 20
<210> 241
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 14
<400> 241
gtcgtccttg aagaagatgg 20
<210> 242
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 15
<400> 242
cggcgcgggt cttgtagttg 20
<210> 243
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 16
<400> 243
gtgtcgccct cgaacttcac 20
<210> 244
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 17
<400> 244
ttcagctcga tgcggttcac 20
<210> 245
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 18
<400> 245
cgtcctcctt gaagtcgatg 20
<210> 246
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 19
<400> 246
agcttgtgcc ccaggatgtt 20
<210> 247
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 20
<400> 247
gtggctgttg tagttgtact 20
<210> 248
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 21
<400> 248
tgtcggccat gatatagacg 20
<210> 249
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 22
<400> 249
accttgatgc cgttcttctg 20
<210> 250
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 23
<400> 250
tgttgtggcg gatcttgaag 20
<210> 251
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 24
<400> 251
agctgcacgc tgccgtcctc 20
<210> 252
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 25
<400> 252
tgttctgctg gtagtggtcg 20
<210> 253
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 26
<400> 253
cacggggccg tcgccgatgg 20
<210> 254
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 27
<400> 254
ctcaggtagt ggttgtcggg 20
<210> 255
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 28
<400> 255
ctttgctcag ggcggactgg 20
<210> 256
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 29
<400> 256
tgatcgcgct tctcgttggg 20
<210> 257
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 30
<400> 257
cacgaactcc agcaggacca 20
<210> 258
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 31
<400> 258
cgagagtgat cccggcggcg 20
<210> 259
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> eGFP smFISH probe 32
<400> 259
cacttgtaca gctcgtccat 20
<210> 260
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 1
<400> 260
gctgcacata tatttatgtc 20
<210> 261
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 2
<400> 261
gcctagaatg cttatgttgt 20
<210> 262
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 3
<400> 262
ggacagtaca catccatttt 20
<210> 263
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 4
<400> 263
atcttcctct ttatacgggg 20
<210> 264
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 5
<400> 264
aggtttcatc agtgtttgga 20
<210> 265
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 6
<400> 265
gaactgcaga gtatgttgtt 20
<210> 266
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 7
<400> 266
attctaggta ggaacgatct 20
<210> 267
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 8
<400> 267
agtcttgttt gcacattctt 20
<210> 268
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 9
<400> 268
catatgcctc atttttccaa 20
<210> 269
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 10
<400> 269
tgaagctcat gactgtgcat 20
<210> 270
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 11
<400> 270
caagtgactt agtgttcccc 20
<210> 271
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 12
<400> 271
accagagttc ctttctcatg 20
<210> 272
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 13
<400> 272
ctctgggaaa ggacaggaca 20
<210> 273
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 14
<400> 273
cttagaagtg ttaggatccc 20
<210> 274
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 15
<400> 274
aactagaggc taccaatggc 20
<210> 275
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 16
<400> 275
ttagcggaca aacagttctg 20
<210> 276
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 17
<400> 276
tcagcatcat tttgctgtta 20
<210> 277
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 18
<400> 277
tctcaacaca catcctgagc 20
<210> 278
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 19
<400> 278
tgacaaaacc cagttctgtc 20
<210> 279
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 20
<400> 279
tgggtgtgaa taacggcttg 20
<210> 280
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 21
<400> 280
caaaccttgg cctaggtgac 20
<210> 281
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 22
<400> 281
cctcatttca cagttgaagt 20
<210> 282
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 23
<400> 282
tttctatggc ctcttgattt 20
<210> 283
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 24
<400> 283
ctcatgcagt aatgctgtga 20
<210> 284
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 25
<400> 284
gccagatatt atgctgtttt 20
<210> 285
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 26
<400> 285
aacaagcttt acctgcagtg 20
<210> 286
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 27
<400> 286
cattttgctt tgagaagtcc 20
<210> 287
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 28
<400> 287
gatgtcctga tattagcaga 20
<210> 288
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 29
<400> 288
agagtctatc ttaagctttt 20
<210> 289
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 30
<400> 289
gattttctca ttactacctt 20
<210> 290
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 31
<400> 290
ctccttctag ctattcttta 20
<210> 291
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 32
<400> 291
ctgattttct cattattacc 20
<210> 292
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 33
<400> 292
ccttctagct atcctttaat 20
<210> 293
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 34
<400> 293
cttgttcttt tcctttcatc 20
<210> 294
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO intron 1 smFISH probe 35
<400> 294
gagcgtgtca cacagagtaa 20
<210> 295
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 1
<400> 295
ggtcttaatc tccacatgta 20
<210> 296
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 2
<400> 296
gagtgttgct gcgagaattt 20
<210> 297
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 3
<400> 297
aagatccaca cataaagcgc 20
<210> 298
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 4
<400> 298
gtagggttaa tatgcagact 20
<210> 299
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 5
<400> 299
tgtttgagag gagccatttc 20
<210> 300
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 6
<400> 300
tacatttggg ctgctctatc 20
<210> 301
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 7
<400> 301
tggaaggcac cacagtagca 20
<210> 302
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 8
<400> 302
tcctcagaga gttttgcatt 20
<210> 303
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 9
<400> 303
ggtagcagta gacatattct 20
<210> 304
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 10
<400> 304
actagatgat gggggttaca 20
<210> 305
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 11
<400> 305
aatgttcacc gtgtaccaac 20
<210> 306
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 12
<400> 306
agcagaatac attcccacag 20
<210> 307
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 13
<400> 307
aattggcagg cctctggaat 20
<210> 308
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 14
<400> 308
ttcagcctat agtgagatta 20
<210> 309
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 15
<400> 309
aagtattcta tgtgagcagc 20
<210> 310
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 16
<400> 310
gtccactgat ctaaactggt 20
<210> 311
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 17
<400> 311
agcagtaatt cagcatctct 20
<210> 312
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 18
<400> 312
ggcatgtgga tctatacttc 20
<210> 313
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 19
<400> 313
actcaacatt cgcctctaat 20
<210> 314
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> UMLILO exon smFISH probe 20
<400> 314
gataaataca gttcctgcct 20
REFERENCES CITED IN THE DESCRIPTION
Patent documents cited in the description
Non-patent literature cited in the description