Anti ci ndufa9

Primary antibodies included anti-acetyl-lysine (Cell Signaling #9441), anti-SIRT3 (Cell Signaling #5490), anti-SOD2 (acetyl K68) (Abcam ab137037), anti-SOD2 (Cell Signaling #13141), anti-VDAC (Cell Signaling #4866), anti-GAPDH (Cell Signaling #5174), anti-FXN (generous gift of Grazia Isaya, Mayo Clinic, Rochester MN), anti-LCAD (Abcam ab196655) and to the electron transport chain complexes: anti-CI-NDUFA9 (Abcam ab14713), anti-CII-SDHB (Abcam ab14714) and anti-CIII-UQCRFS1 (Abcam ab14746). Band intensities were measured using ImageJ (IJ1.46). To determine correlation between acetylation and heart function, relative density of acetylation for n = 3 western blot images was normalized to day 30 controls and averages for each group were used for correlation calculation.

Fibroblasts cell pellets were processed for the analysis of mitochondrial complexes by Blue Native (BN) PAGE as described (Nijtmans et al. 2002 (link)). Part of these mitochondrial protein complexes were further processed for SDS-PAGE by adding SDS-PAGE sample buffer. Protein concentrations were measured using a Micro BCA protein assay kit (Thermo Scientific). Mitochondrial extracts were separated on 5–15% BN-PAGE gels (Nijtmans et al. 2002 (link)), or on 10% SDS-PAGE.
Antisera used were anti-V5 (Invitrogen), anti-COX-I (Abcam), anti-COX-II (Invitrogen), anti-COX-IV (Abcam), anti-Porin/VDAC (Mitosciences), anti-CI (NDUFA9) (Abcam), anti-CII (SDHA) (Abcam), and anti-CIII (UCCRC2) (Abcam).