Fig. 4

ARTADIs are unique compared to enzalutamide in blocking the cell cycle and DNA-damage repair pathways in AR-V7-dependent cells. a Heatmap displaying normalized enrichment scores (NES) using gene set enrichment analysis (GSEA) on RNA-seq data from androgen-deprived LNCaP95 cells treated with enzalutamide (ENZA, 5 µM), ralaniten (EPI-002, 35 µM), BU3-12 (5 µM), or DMSO (VEH) for 48 h. Gene sets were restricted to MSigDB set H (hallmark gene sets). Gene sets with nominal p < 0.05 and false discovery rate (FDR) q < 0.05 were considered significant. b Enrichment plots from selected gene sets showing enriched genes in ENZA treated samples compared to ARTADI (EPI-002, BU3-12). c Cell cycle distribution of LNCaP95 cells labeled with BrdU-FITC and 7-AAD after treatment with DMSO (VEH), enzalutamide (ENZA, 5 µM), ralaniten (EPI-002, 35 µM), masofaniten (EPI-7386, 5 µM), BU130 (5 µM), BU170 (10 µM), or BU3-12 (5 µM) determined by flow cytometry. d Percentage of γH2AX positive LNCaP95 cells after same treatment as in part c. For (c, d), bars represent the mean ± SEM of 3 independent experiments analyzed with two-way ANOVA and Tukey correction. e Transcript levels of AR and cell cycle related genes normalized to housekeeping gene SDHA. LNCaP95 cells were treated with DMSO (VEH), enzalutamide (ENZA, 5 µM), ralaniten (EPI-002, 35 µM), masofaniten (EPI-7386, 5 µM), BU130 (5 µM), BU170 (10 µM), or BU3-12 (5 µM) for 48 h in media supplemented with 1.5% CSS. Data presented as mean ± SEM and normalized to DMSO vehicle and were analyzed by one-way ANOVA with Dunnett’s correction (n = 3 independent experiments). *p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001. See also Supplementary Figs. 2 and 3