Succinate dehydrogenase

Nrf2 (Active Motif, Carlsbad, CA), antioxidant enzymes and (CAT, GPx, and SOD) and aconitase (Cayman Chemical, Ann Arbor, MI) activities, and other mitochondrial activities (Succinate Dehydrogenase, and NADH Dehydrogenase) (Abcam, Cambridge, MA) were measured using commercially available kits as previously described [32] (link). Nrf2 binding was measured using this kit to best assess its binding to the electrophile response element (EpRE). Nuclear fractions were prepared for Nrf2 activity using a NE-PER Nuclear cytosolic extraction kit (Thermo Fisher Scientific, Waltham, MA). Nuclear purity was cross probed with GAPDH and H3 (Cell Signaling, Danvers, MA). Mitochondrial fractions were obtained according to the aconitase assay instructions. Mitochondrial purity was cross probed using GAPDH and VDAC1 (Abcam, Cambridge, MA). All samples were analyzed in duplicate and run in a single assay with intra-assay and percent coefficients of variability of less than 10% for all assays.