Characterization and suitability of the HEMn cell system: The OCA2 gene is differentially expressed in HEMn-LP and HEMn-DP cells. (A) UCSC Browser NCBI36/hg18 assembly (http://genome.ucsc.edu/cgi-bin/hgGateway?db=hg18) overview of the OCA2-HERC2 locus (top panel). (Middle panel) The region covered on BAC RP11-1365A12. Vertebrate conservation (green); the position of rs12913832 (red). (Lower panel) A schematic overview of the region investigated in this study. Restriction enzyme digestion sites are indicated. (B) Sequence analysis of the region around HERC2 rs12913832 in HEMn-LP (left) and HEMn-DP (right). The genotypes of rs12913832 were determined by direct sequencing of PCR fragments containing rs12913832. (C) RT-qPCR analysis of OCA2 primary transcripts in MCF7 and HEMn cells demonstrates differential OCA2 expression between HEMn-LP and HEMn-DP cells. Each gene expression analysis is carried out in triplicate and normalized to an endogenous reference gene (ACTB). (D) ChIP-qPCR of RNA Pol II binding at the OCA2 promoter in MCF7, HEMn-LP, and HEMn-DP cells. Enrichment is calculated relative to necdin (NDN), and values are normalized to input measurements. All ChIP analyses are performed in triplicate. Data are represented as mean ± SEM; (*) p < 0.05; (**) p < 0.01.
