Western Blot Analysis of Protein Markers

Western blotting was performed as described previously [25 (link)]. Briefly, for Western blotting, proteins were extracted using RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% sodium deoxycholate, 0.1% SDS, 1mM Na2EDTA, 1% Triton X-100, 1 μg ml^-1 leupeptin) supplemented with protease inhibitors and phosphatase inhibitors (Biotool). The lysate was centrifuged at 13000 rpm at 4°C for 15 min. The supernatants were collected and protein concentrations were measured using the BCA method. Proteins were then immunoblotted on the PVDF membrane by standard procedures. Antibodies against RIF1 were obtained from Bethyl Laboratories. Antibodies to Bax, Bcl2, N-Cadherin, MMP2, and Integrin-β1 were purchased from Cell Signaling Technology. Antibodies against caspase3 were obtained from Santa Cruz. Antibody to β-actin was purchased from Sigma-Aldrich and used as a loading control.