Fig 1: Processed intracellular domain of JAG1 (JICD) increases the expression of AR-Vs. (A) Representative Western blot analysis showing the increased protein levels of AR and AR–Vs in AdJICD–infected LNCaP and CWR22Rv1 cells compared to AdCtrl–infected cells. The proteins of AR and ARs including AR–V7 were detected using mouse anti-AR (sc–7305, Santa Cruz) and rabbit anti-AR–V7 (68492, Cell Signaling, Danvers, MA, USA) antibodies. (B,C) Stable JICD–overexpressing CWR22Rv1 subline cells show the increased expression of ARs, including AR–V7, at protein (B) and mRNA (C) levels. Representative RT–PCR (top) and quantitative RT–PCR (qPCR; bottom) analysis of three independent experiments (C) revealing a significant increase of AR–V7 mRNA expression in stable JICD–overexpressing CWR22Rv1 subline cells. GAPDH was used as a loading control. Data are shown as mean ± SEM, three independent experiments. *, p < 0.01; ns, not significant; two–tailed t-test. (D) JICD overexpression enriches the expression of genes involved in RNA processing and metabolic process, RNP complex biogenesis, and DNA metabolic process. (E) JICD upregulates the expression of some splicing factors involved in the generation of AR–V7 in CWR22Rv1 cells. (F–H) JICD overexpression enhances the expression of some shared AR/AR–V7 (F) and AR–V7 target genes (G) and negatively affects the expression of most AR target genes (H). False discovery rate (FDR) < 0.05. Values above Western blots indicate the relative band intensity of each protein normalized to GAPDH and are shown as mean ± SD, which were obtained by densitometrical quantification of at least three independent experiments. All uncropped Western blot and gel original images are shown in Figure S7.