Ab13834

After treatment, whole cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Solarbio, R0010) at 4°C for 10 min, and the total proteins were quantified using a bicinchoninic acid (BCA) protein assay kit (Beyotime, P0010S). The proteins were separated via SDS-PAGE and transferred to polyvinylidene fluoride (PVDF, Millipore) membranes. After blocking with 5% skimmed milk, the membranes were incubated with primary antibodies overnight at 4°C. The primary antibodies included monoclonal anti-FLAG M2 (Sigma–Aldrich, F1804, 1:5,000), monoclonal anti-HA tag (Abcam, ab13834, 1:5000), anti-BAX (Abcam, ab53154, 1:5,000), anti-BclXL (ProteinTech, 66020-1-Ig, 1:2,000), anti-Caspase 9 (ProteinTech, 10380-1-AP, 1:1,000), anti-PERK (Abcam, ab65142, 1:500), anti-phosphate eIF2α (Abcam, ab214434, 1:1,000), anti-CHOP (Abcam, ab11419, 1:2,000), anti-Caspase 3 (Abcam, ab208161, 1:1,000), anti-PARP1 (Invitrogen, 436400, 1:2,000) and rabbit polyclonal anti-GAPDH (ProteinTech, 60004-1-Ig, 1:2,000). Following three washes, the membranes were probed with horseradish peroxidase-coupled secondary antibodies. Finally, the membranes were exposed, and signals were recorded using Image Lab software (Bio-Rad) after incubation with enhanced chemiluminescence (ECL) reagents (Beyotime, P0018FM). The relative band intensities were quantified using ImageJ software (V 1.8).

After treatment, whole cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Solarbio, R0010) at 4°C for 10 min, and the total proteins were quantified using a bicinchoninic acid (BCA) protein assay kit (Beyotime, P0010S). The proteins were separated via SDS-PAGE and transferred to polyvinylidene fluoride (PVDF, Millipore) membranes. After blocking with 5% skimmed milk, the membranes were incubated with primary antibodies overnight at 4°C. The primary antibodies included monoclonal anti-FLAG M2 (Sigma–Aldrich, F1804, 1:5,000), monoclonal anti-HA tag (Abcam, ab13834, 1:5000), anti-BAX (Abcam, ab53154, 1:5,000), anti-BclXL (ProteinTech, 66020-1-Ig, 1:2,000), anti-Caspase 9 (ProteinTech, 10380-1-AP, 1:1,000), anti-PERK (Abcam, ab65142, 1:500), anti-phosphate eIF2α (Abcam, ab214434, 1:1,000), anti-CHOP (Abcam, ab11419, 1:2,000), anti-Caspase 3 (Abcam, ab208161, 1:1,000), anti-PARP1 (Invitrogen, 436400, 1:2,000) and rabbit polyclonal anti-GAPDH (ProteinTech, 60004-1-Ig, 1:2,000). Following three washes, the membranes were probed with horseradish peroxidase-coupled secondary antibodies. Finally, the membranes were exposed, and signals were recorded using Image Lab software (Bio-Rad) after incubation with enhanced chemiluminescence (ECL) reagents (Beyotime, P0018FM). The relative band intensities were quantified using ImageJ software (V 1.8).