ChIP Analysis of ER, CBP Targets

ChIP experiments were performed, as previously described.^{32 (link)} Briefly, cells were synchronised with α-amanitin,^{33 (link)} then treated for 45 min with E2 (1 nM), ICI (10 nM), abiraterone (7.5 μM) or combination and fixed. The antibodies used were anti-ER (Santa Cruz Biotechnology; sc-543×), anti-CBP (Santa Cruz Biotechnology; sc-369×) and Mouse IgG1 (Dako, Denmark A/S). The resulting DNA was subjected to quantitative PCR analysis using SYBR green (Applied Biosystems) with the following primers for TFF1: (forward) 5′-GGC CAT CTC TCA CTA TGA ATC ACT TCT GCA-3′ and (reverse) 5′-GGC AGG CTC TGT TTG CTT AAA GAG CGT TAG-3′, and for GREB1: (forward) 5′-GAA GGG CAG AGC TGA TAA CG 3′ and (reverse) 5′-GAC CCA GTT GCC ACA CTT TT.