Fig. 2: ZIC5 regulates EMT progression in PCa cell lines.
From: AR-regulated ZIC5 contributes to the aggressiveness of prostate cancer
A Analysis of the correlation between ZIC5, CDH2, and TWIST1 expression in PCa patients (TCGA-PCa data from ENCORI). B, C Analysis of N-cadherin, Snail1, E-cadherin, and TWIST1 expression in C4-2B and 22RV1 cells transfected with ZIC5-targeted shRNA (sh-ZIC5) or ZIC5 overexpression plasmid (oe-ZIC5), measured by western blotting (B) or RT-qPCR (C). *P < 0.05, relative to sh-NC or oe-NC. D Putative ZIC5-binding sites on the TWIST1 promoter region. E, F ZIC5-targeted shRNA or ZIC5 overexpression plasmid and TWIST1 promoter-driven wild-type luciferase reporter or mutant vectors were cotransfected into C4-2B (E) and 22RV1 (F) cells. Luciferase assays were performed to examine ZIC5/TWIST1 interaction. *P < 0.05, relative to sh-NC or oe-NC. G ChIP-qPCR analysis of ZIC5 binding to the promoter region of TWIST1. C4-2B cells were transfected with Flag-tagged ZIC5 or control vectors, followed by immunoprecipitation with anti-Flag or anti-IgG antibodies. Purified IgG was used as control. Experiments were performed in triplicate. *P < 0.05 vs. IgG. H Representative ChIP results showing ZIC5 recruitment onto the TWIST1 promoter.
