Fig. 5

ARTADIs are unique compared to enzalutamide in blocking the cell cycle and DNA-damage repair pathways in cells that are dependent on full-length AR. a Heatmap of normalized signal intensity following RNA-seq transcriptional profiling of LNCaP cells pre-treated with 5 μM enzalutamide (ENZA), 35 μM EPI-002 (ralaniten), 5 μM BU3-12 or DMSO (VEH) for 16 h and stimulated with 1 nM R1881 (+) for 24 h from n = 3 biological replicates. b PCA plot based upon transcript counts for 3793 genes. c Volcano plot depicting androgen-regulated genes defined as genes which with a 2-fold change in expression (VEH+ vs VEH-) and FDR < 0.01. 1094 and 821 genes were defined as androgen-induced and androgen-repressed respectively. d Venn diagrams showing the ability of EPI-002, BU3-12 and/or enzalutamide to repress androgen-induced genes (left) or derepress androgen-repressed genes 2-fold or greater. 168 androgen-induced genes and 49 androgen-repressed genes were not affected by any treatment. e Heatmap showing normalized enrichment scores (NES) from GSEA. Gene sets were restricted to MSigDB set H (hallmark gene sets). Gene sets with nominal p < 0.05 and FDR q < 0.05 were considered significant. f Enrichment plots from selected gene sets comparing all treatments versus vehicle plus androgen (VEH + , top left) or ARTADI inhibitors EPI-002 and BU3-12 versus enzalutamide (top right and bottom). g Cell cycle distribution of LNCaP cells exposed for 48 h to DMSO, enzalutamide (ENZA, 10 µM), EPI-002 (35 µM), EPI-7386 (10 µM), BU130 (5 µM), BU170 (10 µM), and BU3-12 (10 µM). h γH2AX-positive LNCaP cells treated as in (g). Data in (g, h) represent the mean ± SEM of 3 independent experiments analyzed with one-way ANOVA test with Tukey correction. **p < 0.01, ****p < 0.0001. See also Supplementary Figs. 4 and 5