Anti n cadherin

Cells or tissues with different treatments were collected and lysed with RIPA buffer containing phosphatase/protease inhibitor to obtain total cellular protein. Equal amounts of proteins (30 µg) were subjected to SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 5% BSA for 2 h and incubated with primary antibodies overnight at 4 °C. Primary antibodies including anti-CitH3 (1:1000, ab5103, Abcam, Cambridge, England, UK), anti-PAD4 (1:1000, GTX113945, geentex, San Antonio, Texas, USA), anti-N-cadherin (1:1000, YM0465, Immunoway, Plano, Texas, USA), anti-E-cadherin (1:1000, 3195S, CST, Boston, Massachusetts, USA), anti-vimentin (1:1000, sc-6260, Santa Cruz, Dallas, Texas, USA), anti-VEGF (1:200, CY5096, Abways, Minhang District, Shanghai, China), anti-MMP9 (1:1000, ab283575, Abcam, Cambridge, England, UK), anti-MMP2 (1:1000, ab92536, Abcam, Cambridge, England, UK), and anti-β-actin (1:1000, Zoonbio Biotechnology, Nanjing, Jiangsu, China), followed by incubation with horseradish-peroxidase-conjugated secondary antibodies for 1 h at 37 °C. The peroxidase activity of secondary antibodies was detected with ECL Western blot substrate and recorded by Bio-Rad Electrophoresis Documentation (ECL, Millipore, Germany).

An equal amount of total protein was run on 12.5% SDS-PAGE, transferred to PVDF membranes (250 mA for 2 h), and probed with primary antibodies. The primary antibodies include anti-MMP2 (1:1000, 72 kDa, Sigma-Aldrich), anti-N-cadherin (1:1000, 140 kDa, ImmunoWay Biotechnology), anti-E-cadherin (1:1000, 135 kDa, ImmunoWay Biotechnology), anti-Vimentin (1:1000, 57 kDa, ImmunoWay Biotechnology), anti-ADAM9 (1:1000, 72 kDa, Affinity Biosciences), anti-Snail (1:1000, 29 kDa, ImmunoWay Biotechnology), anti-DNMT1 (1:1000, 183 kDa, Abcam), anti-CD47 (1:1000, 52 kDa, Abcam), and anti-GAPDH (1:1000, 37 kDa, Sigma-Aldrich). The protein bands of interest were captured after the secondary antibodies linked with peroxidase were bound to primary antibodies [26 (link)].