Quantification of 5 electron transport chain complexes using homogenized soleus muscle reveals no significant differences in the PAS kinase-deficient male (PASK−/−) mice compared to the WT. (A) Soleus tissue was homogenized and analyzed by western blot using the OxPhosBlue Native WB Antibody Cocktail (ThermoFisher Scientific, Waltham, MA, USA) containing mouse monoclonal NDUFA9 (complex I), SDHA (complex II), UQCRC2 (complex III, core II), COX IV (complex IV, subunit IV) and ATP5A (complex V alpha subunit) antibodies. Protein concentration was determined before loading using the Pierce Coomassie Plus (Bradford) Assay Reagent (ThermoFisher Scientific, Waltham, MA, USA). The same control sample (cntrl) was loaded on each gel for normalization between gels. (B) Plots of one-factor (genotype or diet) analysis of complex I. (C) Representative western blots for each complex are shown. Each biological replicate n > 4 was run in duplicate. Error bars represent SEM. Two-factor (genotype and diet) ANOVA was performed using JMP Pro14 software with students t-test performed on significant differences, * p < 0.05 is shown.