Western Blot Analysis of EMT Markers

Western blot analyses were performed as described previously^{43 (link)}. Samples, which were lysed in buffer containing 150 mM NaCl, 10 mM Tris-HCl (pH 7.4), 2.5 mM EDTA, 0.125% Nonidet P-40 (v/v), and protease inhibitors, were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Bedford, MA, USA). Membranes were probed with anti-β-catenin (610154; BD Biosciences, San Jose, CA, USA), anti-TGF-β2 (ab36495; Abcam, Cambridge, MA, USA), anti-E-cadherin (610181; BD Biosciences, San Jose, CA, USA), anti-vimentin (SC-6260; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-actin (SC-1615; Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibodies. Immunoreactivity was visualized by autoradiography.