Fig. 4

Over-expressed androgen receptor competes with ERα for AIB1 interaction. a MCF-7 cells, were transiently co-transfected with pCD1prom-Luc (0.25 μg /well) and /or pcDNA3-AR (AR) and/or increasing amounts (given in μg/well) of full-length AIB1 expression plasmid (AIB1), and treated as indicated. Columns are mean of three independent experiments and expressed as fold induction over untreated, which was assumed to be 100%; bars SD; *p ≤ 0.05. b Total cell extracts from MCF-7 and MCF-7/AR were immunoprecipitated (IP) with an anti-AIB1 antibody and immunoblotted (WB) to detect AIB1, ER and AR protein levels. Results are representative of three independent experiments. c Total cell extracts from Hela cells transiently cotransfected with different amounts (given in μg/dish) of pcDNA3-AR (AR), Hego (ER) or full-length AIB1 expression plasmid as indicated were immunoprecipitated (IP) with an anti-AIB1 antibody and immunoblotted (WB) to detect AIB1, ER and AR protein levels. Results are representative of three independent experiments. d and e ChIP-qPCR performed on MCF-7 cells and MCF-7/AR cells using anti-AIB1 (d) or anti- ERα (e) antibodies, as indicated. IgG was used as control. Columns are the mean of three independent experiments. Bars, SD; *p ≤ 0.05 vs untreated MCF-7 cells; ^□p ≤ 0.05 vs. E₂-treated MCF-7 cells; ^●p ≤ 0.05 vs E₂ + DHT treated MCF-7 cells