Cells were lysed using radioimmunoprecipitation assay buffer (Beyotime, China) and then quantitatively assessed using a bicinchoninic acid protein assay kit (Pierce, USA) and NanoDrop ND 2000 c Spectrophotometer (Thermo Scientific, USA) as per the manufacturer’s instructions. Equal amounts of denatured proteins were loaded onto 10% sulfate-polyacrylamide gel and separated via electrophoresis run at 75 to 175 V. The proteins in the gels were transferred onto polyvinylidene fluoride membranes using a wet transfer system and blocked with 5% milk in TBS at 25°C for 1 h. Membranes were continuously exposed to TBST containing anti-CKS1B antibodies (Cat#:C0748, Sigma-Aldrich, USA), anti-Bax antibodies (Cat #33-6400, 1:1000, Thermo Fisher Scientific, USA), anti-Bcl-2 antibodies (Cat #MA5-11,757, 1:1000, Thermo Fisher Scientific, USA), or anti-GAPDH antibodies (Cat #MA1-16,757,1:1000, Thermo Fisher Scientific, USA) at 4°C. Antibody-bound proteins were probed using an anti-mouse/goat HRP secondary antibody (Cat #31,430 or Cat #31,402, 1:1000, Invitrogen, USA) at room temperature for 1 h. Chemiluminescent detection (Beyotime, China) was used to visualize the target proteins, and the intensity of protein signals was estimated using Quantity One 4.1 (Bio-Rad Laboratories, USA) [22 (link)].