Fig. 3: UBE2J1-loss leads to induction of AR and AR signaling.

A GSEA Pathways analysis shows cancer-related signaling pathways significantly altered in UBE2J1-KO cells compared to wildtype cells, AR related and luminal lineage pathways were highlighted in orange and lineage specific pathways were highlighted in red. Reads from three biological replicates were used for analysis. B GSEA analysis represents the expression of hallmark androgen response gene signature in UBE2J1-KO cells compared to wild type. Reads from three biological replicates were used for analysis. C Relative gene expression of canonical AR target genes, measured by qPCR, in LNCaP/AR cells transduced with Cas9 and annotated guide RNAs, then treated with CSS + 10 µM ENZ for 5 days. n = 3 independently treated cultures, mean ± s.e.m. is presented. p values were calculated using two-way ANOVA with Bonferonni multiple-comparison test. D Western blots represent AR and Actin proteins in LNCaP/AR cells transduced with Cas9 and annotated guide RNAs, then treated with Veh (DMSO) in full serum or CSS + 10 µM ENZ for 5 days. E Western Blot represents AR-V7 and Actin proteins in LNCaP/AR cells transduced with Cas9 and annotated guide RNAs under Veh (DMSO). F Western Blot represents AR and Actin proteins in LNCaP/AR cells transduced with Cas9 and annotated guide RNAs, then treated with Veh (−DHT) or 10 nM dihydrotestosterone (DHT) for 48 h. G, H Representative immunofluorescence staining of LNCaP/AR cells transduced with Cas9 and annotated guide RNAs with annotated antibodies, scale bar represents 50 µm. Cells were treated with G 10 µM ENZ or H Veh (DMSO) for 7 days. I Immunohistochemical staining of AR and NKX3.1 proteins on sgNT and sgUBE2J1 xenograft tumor slides from castrated and ENZ treated mice, scale bar represents 50 µm. J Immunohistochemical staining of AR and NKX3.1 proteins on sgNT and sgUBE2J1 xenograft tumor slides from intact and Veh treated mice, scale bar represents 50 µm. For panels G, H, statistical analysis of representative pictures (n = 4–6) is presented, and p values were calculated using two-tailed t-test. For all panels, mean ± s.e.m. is presented.