Gene Regulatory Scenarios of Primary 1,25-Dihydroxyvitamin D3 Target Genes in a Human Myeloid Leukemia Cell Line
Abstract
:1. Introduction
2. Experimental
2.1. Cell Culture
2.2. RNA Extraction, cDNA Synthesis and qPCR
2.3. ChIP
3. Results
3.1. Transcription of G0S2, CDKN1A and MYC in Monocyte- and Macrophage-Like Cells
3.2. Genomic Profile of G0S2, CDKN1A and MYC in Monocyte- and Macrophage-like Cells
3.3. VDR Binding Sites of G0S2, CDKN1A and MYC
4. Discussion
5. Conclusions
Acknowledgments
Conflicts of Interest
References
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Supplementary Materials
Gene | Fragment size (bp) | Annealing temperature (°C) | Primer sequences (5'-3') |
---|---|---|---|
B2M 1,2 | 246 | 60 | GGCTATCCAGCGTACTCCAAA
CGGCAGGCATACTCATCTTTTT |
CDKN1A 3 | 99 | 60 | GCCACTAAGGTCATTCCCGCCT
CCTTGCGCTTCTGGGCCATCAT |
G0S2 4 | 102 | 60 | GCCACTAAGGTCATTCCCGCCT
CCTTGCGCTTCTGGGCCATCAT |
GAPDH 1,2 | 113 | 60 | CATGAGAAGTATGACAACAGCCTAGTC
CTTCCACGATACCAAAGT |
HPRTI 1,5 | 94 | 60 | TGACACTGGCAAAACAATGCA
GGTCCTTTTCACCAGCAAGCT |
MYC 3 | 147 | 60 | CCAGCAGCGACTCTGAGG
GGACCAGTGGGCTGTGAG |
Genomic region | Fragment size (bp) | Annealing temperature (°C) | Primer sequences (5'-3') |
---|---|---|---|
G0S2 1 | 119 | 62 | GCCTGAGAGTATGCTGTGTACGTTT CTAAGTGCTCTCTTGGCGTAACAAA |
CDKN1A 2, peak 1 | 129 | 58 | TGTGGGGAGGGTGTTTCAG GAGGGAAGGAAGGAGTGAG |
CDKN1A 2, peak 2 | 91 | 58 | CAGAGGAAGTGGGTTGAG AGCAGGGCAGGAGAGATTATAC |
CDKN1A 2, peak 3 | 69 | 62 | GCACTCTTGACCTTGACGGA CTAACACCCTTGGCTTGGAC |
CDKN1A 2, control | 180 | 62 | ATCACAGGGGTCAGCACATC CGCAGCATTTGGGTTCACAC |
MYC 2, peak 1 | 181 | 62 | GTCACACCCTTCTCCCTTC CGCTCCACATACAGTCCTG |
MYC 2, peak 2 | 93 | 58 | CCACACTAACCTCTCAGTTC GTAATGATACTCCCAGCAAAG |
MYC 2, peak 3 | 137 | 62 | GGATGTCAGCAGGGTTTCTC GGAAGTGATTTCGGGAGTAG |
MYC 2, peak 4 | 96 | 58 | GCTCTGTTGGTGTGGACTG GATTAGGGTGCCATAGAATAC |
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Ryynänen, J.; Seuter, S.; Campbell, M.J.; Carlberg, C. Gene Regulatory Scenarios of Primary 1,25-Dihydroxyvitamin D3 Target Genes in a Human Myeloid Leukemia Cell Line. Cancers 2013, 5, 1221-1241. https://doi.org/10.3390/cancers5041221
Ryynänen J, Seuter S, Campbell MJ, Carlberg C. Gene Regulatory Scenarios of Primary 1,25-Dihydroxyvitamin D3 Target Genes in a Human Myeloid Leukemia Cell Line. Cancers. 2013; 5(4):1221-1241. https://doi.org/10.3390/cancers5041221
Chicago/Turabian StyleRyynänen, Jussi, Sabine Seuter, Moray J. Campbell, and Carsten Carlberg. 2013. "Gene Regulatory Scenarios of Primary 1,25-Dihydroxyvitamin D3 Target Genes in a Human Myeloid Leukemia Cell Line" Cancers 5, no. 4: 1221-1241. https://doi.org/10.3390/cancers5041221