Anti ndufa9

Blue native gel electrophoresis (PAGE) was performed by isolating mitochondrial proteins from MT and WT zebrafish, as detailed previously (50 (link)). Samples containing 15 μg of proteins were separated on 3–11% Bis-Tris Native PAGE gel. The primary antibodies applied for this experiment were anti-Ndufa9 (Proteintech, 20312-1-AP), anti-Sdha (Proteintech, 14865-1-AP), anti-Uqcrc2 (Abcam, ab203832), anti-Cox5a (Proteintech, 11448-1-AP), anti-Atp5c (Proteintech 60284-1-Ig) and anti-Vdac1/2(Proteintech, 10866-1-AP). Peroxidase AffiniPure goat anti-mouse IgG and goat anti-rabbit IgG (Jackson) were used as secondary antibodies and protein signals were detected using the ECL system (CWBIO).

Cells were lysed in RIPA lysis buffer (#R0278, Sigma) with the addition of the Halt Protease Inhibitor Cocktail (#8778, Thermo Scientific) and Halt Phosphatase Inhibitor Cocktail and EDTA solution (#78420, Thermo Scientific). The antibodies used included anti-NDUFA9 (#20312-1-AP, Proteintech), anti-COX4 (#GTX114330, GeneTex), anti-COX2 (#55070-1-AP, Proteintech), anti-β-actin (#MAB1501, Millipore), peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) (#115-035-003, Jackson ImmunoResearch), and peroxidase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (#111-035-003, Jackson ImmunoResearch). The signals were developed using the Ultra ECL-HRP Substrate (#TU-ECL02, TOOLS) using X-ray films.