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).