Fig. 1

Cox7a1 stabilizes the dimerization of complex IV (A) BNGE and immunoblotting with the indicated antibodies of cox7a1+/+ and cox7a1−/− of whole zebrafish mitochondria. Shown are representative examples from a total of 3 technical replicates. Arrowheads mark regions where CIV complexes are missing. VDAC was used as loading control. (B and C) BNGE plot profile from CIV and CIII immunodetection with the indicated antibodies. (D) 2D BNGE digitonin-DDM and immunoblotting with the indicated antibodies of whole zebrafish mitochondria. Shown are representative examples from a total of 2 technical replicates. Arrowheads, missing CIV staining. (E) BNGE of adult cox7a1+/+ and cox7a1−/− whole body mitochondria (pool of n = 5 fish) and 48 hours post fertilization (hpf) deyolked wild-type embryos (pool of approximately 500 embryos; n = 1 technical replicate). Arrowheads, missing CIV complex. (F) BNGE band oxygen consumption rate normalized by complex IV sample content measured by MT-CO1 immunoblotting of CIV dimer (CIVd or CIV2) and monomer (CIVm or CIV1) of C57BL/6 mice heart mitochondria (n = 5 biological and technical replicates). Unpaired t test. (G) CIV enzymatic activity normalized by citrate synthase activity in skeletal muscle zebrafish mitochondria (n = 5 biological and technical replicates). Unpaired t test. (H) Oxygraphy of skeletal muscle mitochondria from zebrafish (n = 8 biological replicates, one technical replicate). Asterisk highlights changes between curves. (I) Oxygraphy of whole-body mitochondria from zebrafish (n = 5 biological and technical replicates). Genotypes were cox7a1−/− and wild-type sibling as explained in the legend. Two-way ANOVA. Ns, non-significant p > 0.05.