Anti snail ab180714

GW4869 was obtained from Sigma-Aldrich (St. Louis, MO, United States). Lipofectamine® 3,000 Transfection Reagent was purchased from Thermo Scientific (USA). RIPA lysis buffer was purchased from Beyotime (Jiangsu, China). Primary antibodies for immunoblot analysis included anti-ADAM17 (ab39163), anti-CD81 (ab79559), anti-E-cadherin (ab231303), anti-N-cadherin (ab76011), anti-vimentin (ab92547), anti-snail (ab180714), and anti-β-actin (ab8226) (Abcam, Cambridge, MA, United States). Goat anti-rabbit IgG and goat anti-mouse IgG antibodies were purchased from LI-COR (Lincoln, NE, United States).

Briefly, HEC-1B cells were seeded on the confocal laser dish at a density of 5 × 10⁴ cells/ml and cultured for 24 h. The cultured cells were washed 3 times with PBS and fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, USA) for 30 min. After blocking, the cells were incubated first with anti-E-cadherin (3195; 1:100; CST), anti-N-cadherin(13116; 1:200; CST), anti-Vimentin (5741; 1:200; CST) or anti-Snail (ab180714; 1:50; abcam) antibody overnight at 4°C, and then with Cy3-conjugated or FITC-conjugated goat anti-rabbit IgG antibody (1:200; Abcam, Cambridge, UK) and 5 mg/ml DAPI (Sigma-Aldrich) at room temperature for 30 min. Then, the cells were observed under a laser scanning confocal microscope (Leica, Heidelberg, Germany) with emission wavelengths of 518 nm, 570 nm, and 461 nm.