Fig. 5

RARγ governs androgen-induced responses in non-malignant HPr1-AR prostate epithelial cells. HPr1-AR cells were stable transfected with shRNA to RARG and the response to DHT measured. a Cell viability of HPr1-AR-shCTL and HPr1-AR-shRARG cells in the absence or presence of 10 nM DHT was measured for up to 96 h. Significance of differences between triplicate experiments between viability of indicated experimental groups at the end of the study is noted. b HPr1-AR-shCTL and HPr1-AR-shRARG cells were treated in triplicate with DHT or vehicle control for 24 h and gene expression profiled via RNA-seq and analyzed with Rsubread/DESeq2. Volcano plot depicting DHT-induced gene expression changes in HPr1-AR-shCTL cells (top). Highlighted genes represent those that display induction (purple) or repression (green) in response to DHT in HPr1-AR-shCTL expression, and which were significantly dampened in HPr1-AR-shRARG cells (bottom); 6 genes are labeled that had the greatest magnitude of dampened response, including a loss in induction of TMEM37 and a loss of repression of FGFBP1. c Venn diagram representing the number of DEGs determined after DHT treatment in HPr1-AR-shCTL and HPr1-AR-shRARG cells. Specifically, 1454 of the 2309 (63.0%, hypergeometric p value < 2e⁻¹⁶) DHT-regulated DEGs were either significantly less induced or had significantly reduced repression in shRARG cells. d Heatmap depicting normalized enrichment scores (NES) of all enriched pathways related to DHT treatment in different comparisons (NES > 1.8, FDR q-value < 0.05). Select meta-groups from keyword enrichment analysis are depicted. e Example of top significantly upregulated (left) and downregulated (right) GSEA pathways upon DHT treatment. Enrichments are shown for each comparison, as well as (f) an expression profile of a representative gene from each gene set (left, TMEM37, right TFB2M)