Figure 1.

High-dose androgen treatment enhances global Rb binding to suppress E2F signaling

(A) C4-2 cells were stably infected by doxycycline-inducible lentiviral shRNA against non-target control (shNTC) or RB1 (shRB, C4-2-tet-shRB). RNA-seq was done in C4-2-tet-shRB cells (no doxycycline treatment) stimulated by ethanol or 10 nM DHT for 24 h (hours). The gene profiling data for VCaP and VCaP-CR cells stimulated by ethanol and DHT (10 nM, 24 h) were obtained from a previous study.² Gene set enrichment analysis (GSEA) was done to compare the androgen-repressed genes in C4-2-tet-shRB versus VCaP/VCaP-CR cells. NES, normalized enrichment score. (B) Enrichments of HALLMARK_E2F_TARGETS and HALLMARK_MYC_TARGET_V1 gene sets were plotted. (C) GSEA was done to compare directly androgen-repressed genes (AR-repressed genes with nearby AR binding) in these cells. (D–G) ChIP-seq analyses were performed in C4-2 cells treated with ethanol or DHT (10 nM, 4 h). Venn diagrams (D and E) or heatmap view (F and G) for Rb or c-Myc binding peaks were shown. (H) Binding and expression target analysis (BETA) for the association of Rb or c-Myc binding sites (DHT stimulated) with the expression of AR-repressed genes identified from RNA-seq (A). n.s., not significant.