Superoxide dismutase

Biomarkers were assayed using commercially available kits in accordance with manufacturer’s guidelines, including fasted non-esterified FAs (NEFA) (Solarbio), lactate dehydrogenase (LDH) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), creatine kinase-MB (CK-MB) (Nanjing Jiancheng Bioengineering Institute), malondialdehyde (MDA) (Beyotime Institute of Biotechnology, Shanghai, China) and superoxide dismutase (SOD) (Beyotime Institute of Biotechnology).

3% H₂O₂ solution and dimethyl sulfoxide were obtained from Sigma (St. Louis, MO). RPMI-1640 culture medium, 0.05%Trypsin-EDTA, and fetal bovine serum (FBS) were from Gibco (Grand Island, NY, USA). One step TUNEL apoptosis assay kit, puromycin dihydrochloride, bicinchoninic acid assay kit, glutathione peroxidase kit, catalase assay kit, DAPI staining solution, DAF-FM diacetate kit, dihydroethidium, superoxide dismutase, and malondialdehyde assay kit were obtained from Beyotime (Shanghai, China). Anti-Bax, anti-Bcl-2, and anti-Caspase-3 were from Abcam (Cambridge, MA, USA). Anti-PI3K, anti-Akt, antiphosphorylation-PI3K, and antiphosphorylation-Akt were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Immobilon-PSQ transfer membrane was obtained from Millipore (Billerica, MA, USA). An Annexin V/FITC apoptosis detection kit was from BD Biosciences (San Diego, CA, USA). PI3-kinase LY 294002 was purchased from MedChemExpress LLC (New Jersey, USA). Lipofectamine™ 3000 transfection reagent was purchased from Thermo Fisher Scientific, Inc. (Invitrogen, USA). Lentivirus control and PI3K shRNA (U6-MCS-Ubiquitin Cherry-IRES-puromycin) were purchased from GeneChem (Shanghai, China).