Extended Data Fig. 5: Activation of AR relocates p300 from ER to AR target genes.

a, Replicate data for p300 ChIP-seq in vitro experiments in ZR-75-1 cells associated with Fig. 2e. Venn diagrams show the overlap of three independent experiments representing consecutive passages of cells treated with vehicle (Veh), estrogen (E2; 10 nM) or estrogen plus androgen (E2 + DHT; 10 nM each). Peaks present in at least 2 of 3 replicates were used to generate a consensus cistrome, indicated below the Venn diagrams, for further comparative analyses. b, Overlap of consensus p300 cistromes under E2 or E2 + DHT hormone treatments, after subtracting peaks present under basal (Veh) conditions to generate as set of hormone-regulated peaks. c, Consensus p300 ChIP-seq data from (b), associated with Fig. 2e, showing average read density plots (top panels) and heatmaps (bottom panels), illustrating changes in hormone-regulated p300 chromatin binding sites following treatment with E2 + DHT. d, Average read density plots for p300 binding at ER binding sites (ERBS) proximal (<100 kb) to genes down-regulated by androgen under estrogenic conditions (left panel), and AR binding sites (ARBS) proximal (<100 kb) to genes up-regulated under the same conditions (right panel). e, Example genome browser images showing averaged p300 ChIP-seq signals at binding sites associated with an ER target gene (PGR; left panel) and AR target gene (SEC14L2; right panel) in ZR-75-1 cells. Data represents the average signal of three replicates. f, p300 ChIP-qPCR at enhancers of ER-regulated cell cycle genes (as per Fig. 2g) in T-47D cells treated in vitro under designated hormone conditions. Data was analyzed by a two-way ANOVA (F = 93.45, 80.19, and 14.78 for hormone treatment (p < 0.0001), test site (p < 0.0001), and their interaction (p < 0.0001), respectively; df = 30). Data represented as mean ± SEM of 3 independent passages of cells. Post-hoc analyzes were performed using Tukey’s multiple comparisons test, where asterisks denote statistical significance; *p = 0.0275; *** p = 0.0040; ****p < 0.0001. g, Heatmap of RT-qPCR data for genes associated with Fig. 2g assessed in ZR-75-1 and T-47D cells treated in vitro with estrogen (E2, 10 nM) alone or in the presence of androgen (E2 + DHT, 10 nM each). Data represents the average normalized gene expression of four experiments conducted on independent passages of cells.