Figure 5. DNA-PKcs regulates a splicing-associated gene signature.

(A) RNA-Seq data derived from CWR22Rv1-AR-EK cells depleted of DNA-PKcs was analyzed for differential splicing activity using SUPPA2 inbuilt statistical test (ref. 65). Events that passed a P value cut off of <0.05 were plotted in the pie chart. (B) Diagrammatic representation and quantification of the statistically significant splicing alterations detected in response to DNA-PKcs depletion, as determined using SUPPA2 inbuilt statistical test (ref 65). (C) Upregulation of AR-V7 in response to darolutamide was validated by Western blotting using an anti-AR-V7 antibody. (D) Differentially expressed splicing-associated genes from DNA-PKcs depleted and NU5455-treated cells were analyzed by GSEA using a darolutamide-responsive gene set (Baumgart et al., ref. 54) to identify splicing factor expression correlating with AR-V7 synthesis. (E) 34 splicing-associated genes were found to be upregulated in response to darolutamide and are shown in the heatmap.