Immunoblotting for Epithelial-Mesenchymal Transition

Immunoblotting was performed as described previously (17 (link)–19 (link)). Protein was extracted from SUM159 cells using 1× RIPA buffer (P0013B, Beyotime, Shanghai, China). Subsequently, the proteins were separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then electro-transferred onto PVDF membranes, which were incubated with primary antibodies overnight at 4°C. The protein bands were detected using enhanced chemiluminescence substrate (P0018M, BeyoECL Plus, Beyotime, China). The antibodies used were as follows: mouse anti-human E-cadherin (1:500, BF0219, Affinit); rabbit anti-human SNAI1 (1:500, 13099-1-AP, Proteintech); rabbit anti-human SMAD3 (1:500, BM3919, BOSTER); rabbit anti-human p-Smad3 (1:500, AP0328, Bioworld); rabbit anti-human N-Cadherin (1:500, No.48495-2), rabbit anti-human METTL3 (1:500, No.31591), and rabbit anti-human GAPDH (1:3000, MB001) all from Bioworld Technology, Ltd).