Figure 7.
FoxA1 regulates CCND1 expression by defining the functionality of enh2 in BCC. MCF7 cells transfected with si LUC or si FoxA1 were used in ChIP experiments analyzing recruitment of ERα (A), p300 (B), and PolII (D) as well as H4 acetylation (AcH4) (C) to the CCND1 regulatory regions. Amounts of immunoprecipitated DNA were normalized to inputs and reported relative to the amount obtained at +8000 bp with si LUC-transfected cells treated with vehicle alone, which was set to 1. Results are means ± SE from two or three independent experiments. (E) Intact nuclei from MCF7 cells transfected with si LUC or si FoxA1 were used in DNase I sensitivity assays performed as in Figure 1. The percent of remaining DNA corresponding to the enh2 (d500 bp) after partial digestion is indicated. (F) DNase I sensitivity assays were performed using primers that spanned ERα-binding sites previously identified (Carroll et al. 2005). p < 0.05 (*) and p < 0.01 (**) versus si LUC-transfected cells. For ER4 and ER13, p values were 0.076 and 0.053, respectively. (G) DNase I sensitivity assays were performed in HeLa, MDA-MB-231, and HepG2 cells as indicated. DNase I sensitivity (+) and insensitivity (−) were determined using the DNase I insensitive control rhodopsin locus as a reference. Results of all DNase I sensitivity assays are from at least two independent experiments.