Pp2a c α

The proteins were extracted from cells treated with 4-HNE by lysis buffer of 2% SDS, cocktail protease inhibitors, and phosphatase inhibitors (1.0 mM Na₃VO₄, 1.0 mM DTT, and 1.0 mM PMSF). The concentration of proteins was measured with BCA Protein Assay kit (Thermo Fisher Scientific Inc., Barrington, IL, USA). Then, these proteins were separated by SDS-PAGE and transferred onto PVDF (Bio-Rad) membranes following the standard procedures. Next, the membranes were blocked by 5% nonfat dry milk in PBST (PBS with 0.1% Tween 20) and incubated with anti-GAPDH, anti-pro-/cleaved-caspase-3, anti-BcL-2, anti-Bax (Billerica, MA, USA), anti-pAKT, anti-AKT, anti-pp70S6K, anti-p70S6K, anti-PP2A-a, anti-PP2A-b′, and PP2A-c-α (Cell Signaling Technology, Beverly, MA, USA) and appropriate secondary antibodies conjugated with horseradish peroxidase and developed with ECL Plus luminescent reagents (Thermo Fisher Scientific Inc., Barrington, IL, USA). The protein level quantification was also carried out by ImageJ.