S1 File - MTO1-Deficient Mouse Model Mirrors the Human Phenotype Showing Complex I Defect and Cardiomyopathy

Figure S1. Survival plot. No differences were detected under standard conditions in the survival of mutant mice (n = 259) as compared to wild-type littermates (n = 359). Figure S2. CI stain, Quantification of complex I in blue-native gels. A, Protein complexes were solubilized with dodecyl-β-d-maltoside, isolated by blue native electrophoresis and stained by in-gel complex I activity assay. Upper panel: muscle, central panel: liver, lower panel brain. B, Complex I was quantified by densitometry (n = 3). +/+, wild-type mice; −/−, Mto1 deletion mutants; BHM, bovine heart mitochondria as ladder; error bars indicate standard deviation (SD). Figure S3. Muscle: Quantification of OXPHOS complexes in muscle tissue. A, Mitochondrial complexes were solubilized with dodecyl-β-d-maltoside and stained with Coomassie. B, Complexes were quantified by densitometry (n = 3). Assignment of complexes: I, complex I; III, complex III; IV, complex IV; V, complex V. +/+, wild-type mice; −/−, Mto1 deletion mutants; BHM, bovine heart mitochondria as ladder; error bars indicate standard deviation (SD). Figure S4. Liver: Quantification of OXPHOS complexes in liver. A, Mitochondrial complexes were solubilized with dodecyl-β-d-maltoside and stained with Coomassie. B, Complexes were quantified by densitometry (n = 3). Assignment of complexes: I, complex I; III, complex III; IV, complex IV; V, complex V. +/+, wild-type mice; −/−, Mto1 deletion mutants; BHM, bovine heart mitochondria as ladder; error bars indicate standard deviation (SD). Figure S5. Brain: Quantification of OXPHOS complexes in tissue from brain. A, Mitochondrial complexes were solubilized with dodecyl-β-d-maltoside and stained with Coomassie. B, Complexes were quantified by densitometry (n = 3). Assignment of complexes: I, complex I; III, complex III; IV, complex IV; V, complex V. +/+, wild-type mice; −/−, Mto1 deletion mutants; BHM, bovine heart mitochondria as ladder; error bars indicate standard deviation (SD). Figure S6. Brain: locomotor activity after paraquat treatment. Reduced locomotor activity of mutants (n = 6) compared to controls (n = 8) after 9 weeks of paraquat treatment (pTable S1. ECG results of an additional cohort of 15 months old mice confirmed the results found in younger animals (cf. Table 1). (PDF)