Figure 2
Selective E 2 -dependent recruitment of NCOA3 to the endogenous PLAC1 promoter in ERα positive MCF-7 cells. (A) Chromatin immunoprecipitation (ChIP) was performed with chromatin prepared from MCF-7 and SK-BR-3 cells which were either untreated (C), treated with 17-β-estradiol (E2) alone, with ICI alone or with both compounds (E2/ICI). The promoter region of PLAC1 containing the C/EBPβ and SP1 elements (−348/-198) and a region upstream of the promoter (−1219/-1064) as negative control were analyzed by PCR following immunoprecipitation with the indicated antibodies. Amplification products from soluble chromatin prior to precipitation are shown as control (Input). (B) Quantitative analysis of the recruitment and occupancy shown in (A) determined by real-time RT-PCR. The results corrected by input are shown as fold increase compared to unstimulated cells used as a reference.