Keap 1 sirna

H₂S donor NaHS, palmitate, MDC, CSE inhibitor PPG, autophagy inhibitor 3MA, disulphide bond inhibitor DTT, mitochondrial ROS inhibitor Mito-TEMPO, ROS inhibitor NAC, E1 inhibitor PYR41 and C-7Az were purchased from Sigma-Aldrich (Sigma, St. Louis, MO, USA). Mito-SOX, DCFH-DA and JC-1 were purchased from Invitrogen (Grand Island, NY, USA). SOD and CAT activity assay kits were purchased from Jiancheng Institute of Bioengineering, Nanjing, China. Keap-1 siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies to cleaved caspase 3 (25546-1-AP), Bax (50599-2-lg), Bcl2 (12789-1-AP), SOD (10269-1-AP), CAT (21260-1-AP), Beclin1 (11306-1-AP), ATG7 (10088-2-AP), P62 (18420-1-AP), LC3 A/B (66139-1-lg), LC3 B (18725-1-AP), Keap-1 (10503-2-AP), ubiquitin (10201-2-AP), VDAC1 (55259-1-AP), GAPDH (10494-1-AP) and β-actin (66009-1-AP) were purchased from Proteintech (Rosemont, IL, USA).

For the RNA interference assay, IL-32- and VHL-specific siRNA oligonucleotides were purchased from Samchully Pharmaceuticals (Seoul, Korea). The following sequences were used for the construction of the siRNAs: IL-32 siRNA forward 5'-GCUCUCUGUCAGAGCUCUU-3' and reverse 5'-AAGAGCUCUGACAGAGAGC-3'; and VHL siRNA forward 5'-CCACAGCUACCGAGUGUAUTT-3' and reverse 5'-AUACACUCGGUAGCUGUGGTT-3'. CHIP siRNA, Keap1 siRNA and PHD2 siRNA were purchased from Santa Cruz Biotechnology. PKCδ siRNA was purchased from Bioneer (Seoul, Korea). The cells were transfected with 10 nM of siRNA using RNAiMAX (Invitrogen, Carlsbad, CA, USA).