Protein Extraction and Western Blot Analysis

Total protein was extracted from the MPC5 cells using RIPA lysis buffer (Beyotime, Jiangsu, China) following the manufacturer's instructions. Western blot analysis was performed as previously described (25 (link)). The antibodies used were the following: rabbit anti-desmin (1:1,000; sc-14026), rabbit anti-snail (1:1,000; sc-28199) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit anti-nephrin (1:2,000; ab136894; Abcam, Cambridge, MA, USA), rabbit anti-Wilms tumor 1 (WT1; 1:500; sc-192), rabbit anti-E-cadherin (1:1,000l; sc-7870) (both from Santa Cruz Biotechnology, Inc.), rabbit anti-GSK3β (1:2,000; ab18893; Abcam), rabbit anti-β-catenin [activated and unphosphorylated, 1:2,000; Netherlands Cancer Institute (NKI), Amsterdam, The Netherlands], mouse anti-β-tubulin (1:3,000; sc-80011), goat anti-rabbit IgG-HRP (1:3,000; sc-2004) and goat anti-mouse IgG-HRP (1:3,000; sc-2005) (all from Santa Cruz Biotechnology, Inc.). For the quantitative analysis of the western blots, the protein band intensities were quantified using Image J software and β-actin was used for normalization.