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Among them, the ERK MAPK pathway is involved mainly in cell proliferation, at the same time, studies have shown that the high activation of ERK is also involved in the process of cell damage and caused cell apoptosis.When PARP-1 activity is disrupted by inhibitors, the amount of activated caspase-3 protein and the number of dead cells are reduced, in addition, JNK1/2 and ERK1/2 protein can be used as the upstream factor of PARP-1 to regulate cell death.DAPI (2-(4-amidinophenyl)-1H-indole-6-carboxamidine) was from Sigma-Aldrich. The Cell Counting Kit-8 (CCK-8) was from Dojindo Laboratories (Tokyo, Japan).The Annexin V-FITC apoptosis detection kit and mitochondrial membrane potential (JC-1) assay kit were purchased from BD Biosciences (San Diego, CA, USA).The dilution of the antibodies were according to the instructions.The NRK-52E cell line was purchased from the Cell Bank of the Institute of Biochemistry and Cell Biology (Shanghai, China), and was used for no more than 15 passages.The NAD/NADH Assay kit was purchased from Suzhou Ered Biological Technology Co. The ATP Assay Kit and redox-sensitive dye DCFH-DA were obtained from Beyotime Biotechnology Co. The scrambled short interfering RNA (si RNA) and PARP-1 si RNAs were synthesized by Invitrogen (Shanghai, China).Rabbit anti-Histone-3H (CST, 9718S), anti-cleaved caspase-3(CST, 9664S), anti-cleaved caspase-9 (CST, 9507), anti-ERK1/2 (CST, 4695S), anti-phosphotyrosine ERK1/2 (CST, 4370S), anti-JNK1/2 (CST, 9252S), anti-phosphotyrosine JNK1/2 (CST, 4668S), anti-p38 (CST, 8690S), anti-phosphotyrosine p38 (CST, 4511S), anti-cyt C (CST, 11940S), anti–COX IV (CST,4890S), anti-β-actin (CST, 4970S) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (Ig G) antibodies were obtained from Cell Signaling Technology Inc. Anti-AIF antibody (abcam, ab110327), anti-Bax antibody (abcam, ab32503), anti-Bcl-2 antibody (abcam, ab136285) were purchased from Abcam Ltd (Cambridge, MA, USA).
The identification and culture of the cells were performed according to previous descriptions95% and the cells were subcultured using 0.25% trypsin-0.02% EDTA digestion.
Anti-PARP-1 antibody (Santa, sc-7150) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).