Fig. 3: AR activity impacts AS landscape distinctively from its transcriptional regulation.

a, b DSEs associated with high and low AR activity (cutoff, Z-score > 7 (blue dashed box) or < −7 (red dashed box)) in pri-PCa (a) and CRPC-Ad (b), respectively. AR activity (see Methods) was used to fractionate patient cohorts followed by splicing analysis by rMATS. Within the violin plots, the center lines represent median values, box edges are 75th and 25th percentiles, and whiskers denote the maximum and minimum values, respectively. c Genomic alterations do not contribute to the diversity of AR activities across PCa populations. Shown are frequency and types of AR mutations observed in TCGA and CRPC cohorts. AR activities of samples were grouped as in a. d, e Overlap between SAGs and DEGs (d), and between SAGs and three sets of AR-regulated genes (e) in indicated contexts. The number in parentheses denotes percentage of overlapped genes  proportional to all SAGs. Circles are not drawn to scale. f–h Experimental design (f), principal component analysis (PCA) showing proper clustering of samples (g), and qPCR validation of intended modulations of AR signaling in LNCaP cells (h). In h, the error bars represent the mean ± SD (n = 9). The P-value was calculated using two-tailed unpaired Student’s t-test. i The AR-regulated AS program in PCa cells. Shown are the DSEs associated with high (up arrow) or low (down arrow) AR activity in LNCaP cells detected by rMATS. j Overlap between SAGs and three sets of AR-regulated genes in indicated contexts. The number in parentheses denotes percentage of overlapped genes  proportional to all SAGs. Circles are not drawn to scale. A3, alternative 3′ splice sites; A5, alternative 5′ splice sites; DEGs, differentially expressed genes; DSEs, differentially spliced events; IR, intron retention; MX, mutually exclusive exons; SAGs, splicing-affected genes; SE, exon skipping.