Rabbit anti bax

Total proteins were extracted from the hippocampus tissues, and protein concentration was assessed by the bicinchoninic acid (BCA) reagent (Thermo Fisher, Waltham, MA, USA) method. Equal amounts of protein were subjected to SDS-polyacrylamide gel electrophoresis. Separated proteins were transferred from the gel onto PVDF membranes (Bio-Rad, Hercules, CA, USA) by a transfer apparatus. The membrane was blocked with 5% fat-free dry milk for 2 h at room temperature. Subsequently, blots were incubated overnight at 4°C with indicated primary antibodies, including rabbit anti-caspase-3 (Sigma, St. Louis, MO, USA, 1 : 1000), rabbit anti-Bax (Sigma, 1 : 1000), rabbit anti-Bcl-2 (Sigma, 1 : 500) and rabbit anti-β-actin (Sigma, 1 : 1000). The membranes were then incubated with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Sigma, 1 : 5000) for 2 h at 37°C. Finally, immunoreactive bands of the protein expression level were normalized to the intensity of the corresponding bands for β-actin. Densitometric analysis of the results was performed with Image J 1.41 software (National Institutes of Health, Bethesda, MD, USA).

Rabbit-anti-TRAF3IP2, rabbit-anti-Cleaved-Caspase 3, rabbit-anti-Caspase 3, rabbit-anti-Bax and rabbit-anti-β-actin antibodies were purchased from Sigma-Aldrich (USA). The TRAF3IP2 siRNA (siTRAF3IP2) and scramble siRNA (siNC) were purchased from Thermo Scientific Open Biosysterms and were cloned into the lentiviral vector GV248 (Jikai, Shanghai, China).