Arhgap24

Samples were treated with RIPA Lysis Buffer (Solarbio, Beijing, China) to extract the total protein. The proteins were quantified and stored at −20°C before use. We prepared 10% sodium dodecyl sulfate polyacrylamide gel to isolate the proteins. After transfer to nitrocellulose membrane, the bands were blocked with 5% non-fat milk. Then, the corresponding primary antibodies and secondary antibodies were diluted to appropriate concentrations and added to the protein bands, respectively. Finally, the protein bands were scanned with Tanon 5200 (Tanon, Shanghai, China). Integrated density value was used to calculate the relative protein quantity. The antibodies and reagents used were: ARHGAP24 (Abcam, ab203874, 1: 500); MMP9 (Abcam, ab76003, 1: 1000); VEGF (Abcam, ab69479, 1: 1000); Vimentin (Cell Signaling Technology, #5471, 1: 1000); E-cadherin (Cell Signaling Technology, #14472, 1: 1000); β-catenin (Abcam, ab32572, 1: 5000); GAPDH (Cell Signaling Technology, #5174, 1: 2000); HRP-labeled Goat Anti-Rabbit IgG (Beyotime, A0208, 1: 1000); HRP-labeled Donkey Anti-Goat IgG (Beyotime, A0181, 1: 1000); HRP-labeled Goat Anti-Mouse IgG (Beyotime, A0216, 1: 1000).