Coactivation of AR and GR synergistically induces a subset of target genes. (A) Aggregate plots represent the eRNA intensity of GRO-seq at intergenic C1 and C2 peaks in EtOH-treated (gray line), Dex-treated (blue line), DHT-treated (red line), and DexDHT-treated (black line) VCaP cells. GRO-seq tags represented in strand-specific manner with separated plus (solid line) and minus (dashed line) strands. Each aggregate plot represents ±1 kb around the center of the binding sites. (B) Venn diagrams depict the overlap of Dex-regulated (blue circle), DHT-regulated (red circle), and DexDHT-regulated (black circle) genes in VCaP cells. (C) Hallmark gene set pathway analysis of synergistic and nonsynergistic genes in VCaP GRO-seq data. Color scale represents −log10 P-value. (D) Association of nonsynergistic (dashed line) and synergistic (solid line) genes with C1 (black color) and C2 (red color) peaks. Data are represented as cumulative distribution function (CDF). Statistical significance calculated with one-way ANOVA with a Bonferroni post hoc test. (E) Example of nonsynergistic FKBP5 and TMPRSS2 genes. (F) Example of synergistic SMIM3, ZBP1, and KRT72 genes. Bar graphs (top) depict target gene expression using GRO-seq tags (TPM values). Genome browser tracks (bottom) depict chromatin binding of AR and GR. Statistical significance calculated with one-way ANOVA with a Bonferroni post hoc test. All genome browser tracks are normalized to a total of 10 million reads. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001.
