Fig. 3.

Biochemical and genetic characterization of mitochondrial phenotype of mpv17 KO larvae. (A) Basal OCR was measured for ∼1 h in fish water, immediately after 4 dpf larvae were exposed to 0.5 μM FCCP and, later, to a combination of 2 μM rotenone (Rot) and 5 μM antimycin (AA). Four independent experiments were performed (n=40). (B) Quantification of basal respiration in wild-type and mpv17^−/− mutants (n=40). (C) Protein blot analysis of different subunits of OXPHOS complexes. (D) Relative quantification of protein amount, using an antibody against βActin for standardization (n=3). R.I., relative intensity. (E) Relative quantification of mtDNA copy number in wild type and mpv17 homozygous mutants. Mean dCt values were calculated as Ct of mt-nd1 (mitochondrially encoded gene) minus Ct of polg (nuclear gene) and plotted with s.e.m. (n=7). (F) Real-time PCR quantification of mRNA transcripts from Ubiquinol-cytochrome c reductase complex subunits (n=6). Statistical analyses were performed using two-tailed Student's t-test. Statistical significance was evaluated by setting a confidence interval of 95%; data are mean±s.e.m. ****P<0.0001; **P<0.01; *P<0.05; ns, not significant. Experiments were performed in biological replicates.