Immunoblot
Cells were lysed with RIPA buffer (150 mmol/L Tris-HCl pH 7.0, 150 mmol/L NaCl, 1% NP-40, 1% Sodium deoxycholate, 0.1% SDS) supplemented with protease inhibitor cocktail (Sigma, p8340) for 30 min at 4oC. Lysates were quantified by Bio-Rad protein assay. The protein samples were then subjected to SDS-PAGE and probed with the indicated antibodies to visualize the protein levels using a chemiScope 6000 Touch (Clinx, Shanghai, China).