Anti snai1 antibody

The frozen tissues (including 6 pairs of OSCC and paired adjacent normal tissues) were dissolved in RIPA buffer containing 1% PMSF (Sigma, St. Louis). 15% SDS-PAGE gels were used for electrophoresis of lysates, and lysates were transferred onto PVDF membranes (Merck Micropore). After blocking with 5% nonfat milk, the membranes were incubated with anti-SNCG antibody (1 : 250; Peking University Cancer Hospital and Institute, China), anti-Snai1 antibody (1 : 500; Cell Signaling Technology, USA), and anti-β-actin antibody (1 : 5000; ZSGB-BIO, China) at 4°C overnight. The membranes were treated with peroxidase-conjugated IgG antibody for 2 hours at room temperature. The protein bands were revealed by detection of enhanced chemiluminescence reagent (Pierce).

Western blot analysis was performed using standard procedures. The following primary antibodies were used in the experiments: anti-FOXM1 antibody (Santa Cruz), anti-pAKT antibody (Peprotech, USA), anti-AKT antibody (Cell Signaling Technology, Beverly, MA, USA), anti-pGSK3β antibody (Proteintech) at 1:1000, anti-Snai1 antibody (Cell Signaling Technology), anti-pIGF1R antibody (Abcam, Cambridge, UK), anti-IGF1R antibody (Abcam, Cambridge, UK), anti-E-cadherin antibody (BD Biosciences, USA), anti-Vimentin antibody (Cell Signaling Technology), anti-N-cadherin antibody (Cell Signaling Technology), anti-pERK1/2 antibody (Abcam), anti-ERK1/2 antibody (Abcam), anti-HIF1α antibody (Novus Biologicals, USA), anti-ETS1 antibody (Abcam) and anti-β-actin antibody (Sigma, St. Louis, MO,USA).