Cytochrome c oxidase 1

Genipin was purchased from Cayman Chemical (Ann Arbor, MI, USA). Cleaved PARP, caspase 3, cleaved caspase 8, caspase 9, Bax, Bim, Noxa, p53 upregulated modulator of apoptosis, Bid, Mcl-1, X-linked inhibitor of apoptosis, Stat3, phospho-Stat3, JAK2, phospho-JAK2, and voltage-dependent anion channel (VDAC), Snai1 antibodies were all purchased from Cell Signaling Technology (Danvers, MA, USA). Bcl-2, B-cell lymphoma extra-large (Bcl-xL), survivin, NADH dehydrogenase (ubiquinone) 1 alpha subcomplex subunit 9 (NDUFA9), succinate dehydrogenase complex flavoprotein subunit A (SDHA), Rieske iron-sulfur (RieskeFeS), cytochrome c oxidase I, and ATP synthase subunit alpha (ATP5A) antibodies were purchased from Santa Cruz Biotechnology. Vimentin was purchased from DAKO (Brüsseler Str. Berlin, German). E-cadherin and N-cadherin were purchased from BD Biosciences (Franklin Lakes, New Jersey, USA). For secondary antibodies, anti-mouse-IgG-horseradish peroxidase (HRP) and anti-rabbit-IgG-HRP were purchased from Cell Signaling Technology. Z-VAD-FMK, a caspase inhibitor, was purchased from Promega (Madison, WI, USA). Ruxolitinib, a JAK2 inhibitor, was purchased from Sigma.

Total proteins from whole kidney tissues and HK2 cells were extracted using a Pro-Prep Protein Extraction Solution (Intron Biotechnology, Gyeonggi-Do, Republic of Korea) according to the manufacturer’s instructions. For Nrf2 expression, nuclear proteins were prepared using the NE-PER nuclear and cytoplasmic extraction kit (Thermo Fisher Scientific, Rockford, IL, USA). Western blot analysis was performed using the following antibodies: Nrf2 (Santa Cruz Biotechnology Inc., Dallas, TX, USA), Keap1 (Santa Cruz Biotechnology Inc.), Lamin B1 (Cell Signaling Technology Inc., Danvers, MA, USA), HO-1 (Cell Signaling Technology Inc.), NQO-1 (Santa Cruz Biotechnology Inc.), SIRT1 (Cell Signaling Technology Inc.), total AMPK (Cell Signaling Technology Inc.), phosphorylated (phospho)-Thr¹⁷² AMPK (Cell Signaling Technology Inc.), PPARα (Abcam), PGC-1α (Novus Biologicals, Littleton, CO, USA), estrogen-related receptor α (ERRα) (Millipore), SOD1 (Enzo Life Sciences, Farmingdale, NY, USA), SOD2 (Abcam), cytochrome c oxidase I (Santa Cruz Biotechnology) and IV (Cell Signaling Technology Inc.), B-cell leukaemia/lymphoma 2 (BCL-2) (Santa Cruz Biotechnology); BCL-2-associaated X protein (BAX) (Santa Cruz Biotechnology) and β-actin (1:10000, Sigma).