Extended Data Fig. 6: Activation of AR relocates SRC-3 from ER to AR target genes; correlating with p300 chromatin localization.

a, Replicate data for SRC-3 ChIP-seq experiments. Venn diagrams show the overlap of three independent experiments representing consecutive passages of ZR-75-1 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 SRC-3 cistromes under E2 or E2 + DHT hormone treatments. c, Two-factor log ratio (M) plot displaying DHT-induced changes in E2-stimulated p300 and SRC-3 enrichment at consensus p300 binding sites in ZR-75-1 cells (as per Fig. 2e). Point color denotes p300 consensus peak occupancy; hormone-responsive E2-unique (pink), hormone-responsive E2 + DHT-unique (purple) and basal (black; plotted at rear). Point co-ordinates denote enrichment scores at consensus binding sites, derived from an average of three replicates. Example p300 binding sites associated with ER (yellow-orange) and AR (orange-red) target genes are highlighted. d, SRC-3 ChIP-seq data showing read density plots (left panel) and genome browser images (right panels), illustrating E2 + DHT-induced changes in SRC-3 enrichment at ER binding sites (ERBS) proximal (<100 kb) to genes down-regulated by androgen under estrogenic conditions. Data represents an average of three replicates. e, Average read density plots for SRC-3 binding at AR binding sites (ARBS) proximal (<100 kb) to genes up-regulated by androgen under estrogenic conditions (left panel). Right panels: Genome browser images displaying averaged SRC-3 ChIP-seq signals at example ARBS; SEC14L2 and ZBTB16. Data represents an average of three replicates. f, SRC-3 ChIP-PCR at enhancers of ER-regulated cell cycle genes (as per Fig. 2g; Extended Data Fig. 5f) in ZR-75-1 cells (upper panel) and T-47D cells (lower panel), treated in vitro under designated hormone conditions. Data was analyzed by a two-way ANOVA. Upper panel F = 235.4, 61.41, and 7.338 for hormone treatment (p < 0.0001), test site (p < 0.0001), and their interaction (p < 0.0001), respectively; df =30. Lower panel F = 33.84, 29.62, and 5.343 for hormone treatment (p < 0.0001), test site (p < 0.0001), and their interaction (p = 0.0003), respectively; df = 30. ChIP-PCR data is 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 < 0.05; ** p < 0.01; *** p < 0.001; ****p < 0.0001. Exact p-values for upper panel: MYC p = 0.0051 for E2 vs. E2 + DHT; BCL2 p = 0.0066 for E2 vs. E2 + DHT; FOXM1 p = 0.296 for E2 vs. E2 + DHT; and p < 0.0001 for all other comparisons indicated by ****. Lower panel: MYB p = 0.0002 for E2 vs. E2 + DHT; CCND1 p = 0.0371 for E2 vs. E2 + DHT; FOXM1 p = 0.0198 for E2 vs. E2 + DHT; p < 0.0001 for all other comparisons that are indicated by ****.