Anti e cadherin 14472

To extract proteins, we utilized radioimmunoprecipitation assay (RIPA) buffer (Beyotime, China, Shanghai) involving protease inhibitor cocktail as well as phosphatase inhibitor (Bimake, Houston, TX, USA), and to identify total protein concentrations, we employed Bio-Rad protein assay (Bio-Rad, Hercules, CA, USA). After polyacrylamide gel electrophoresis (PAGE), the proteins were transferred to polyvinylidene fluoride (PVDF) membrane before being incubated with a specific primary antibody overnight at 4 ℃. Anti-EHD2 (ab222888), anti-Vimentin (ab92547), and anti-GAPDH (ab8245) were supplied by Abcam (Cambridge, MA, USA). Both anti-E-cadherin [14472] as well as anti-N-cadherin [13116] were provided by Cell Signaling Technology (Boston, MA, USA). Both goat anti-rabbit IgG-HRP (abs20040ss) as well as goat anti-mouse IgG-HRP (abs20039ss) were supplied by AiBiXin Biotechnology Co., Ltd. (Absin, China). The PVDF membrane was incubated with the secondary antibody for 1 hour at room temperature. The membrane was observed using ChemiDoc MP Imaging System from Bio-Rad, USA. We used ImageJ (National Institutes of Health, Bethesda, MD, USA) to analyze band intensity.