Fig 2: Analysis of complex I subunits encoded by mitochondrial and nuclear genes. A Western blot analysis of mtDNA encoding proteins. Twenty µg of total cellular proteins from various cell lines were electrophoresed through a denaturing polyacrylamide gel, electroblotted, and hybridized with ND6, ND4L, ND1, ND3 and ND5 antibodies, with ß-actin as a loading control. B Quantification of ND6, ND4L, ND1, ND3 and ND5 in C, CV0, CVND6, M, MV0, and MVND6 cell lines. The calculations were based on three independent determinations in each cell line. The error bars indicate standard error of the mean (SEM). P indicates significance based on Student’s t-test of the differences between M and MVND6 cell lines. C Western blot analysis of nucleus-encoding complex I subunits. Twenty µg of total cellular proteins from various cell lines were electrophoresed through a denaturing polyacrylamide gel, electroblotted, and hybridized with NDUFA10, NDUFS5, NDUFC2, NDUFA11, NDUFS2, NDUFS1 and NDUFB8 antibodies, with ß-actin as a loading control

Fig 3: Analysis of OXPHOS complexes. A In-gel activity of complexes I, II and IV. The activities of OXPHOS complexes from various cell lines after BN-PAGE were measured in the presence of specific substrates [NADH and NTB for complex I, sodium succinate, phenazine methosulfate, and NTB for complex II, DAB and cytochrome c for complex IV]. B Quantification of in-gel activities of complexes I and IV. The calculations were based on three independent determinations in each cell line. C The levels of complexes I, II and IV by BN-PAGE. Twenty micrograms of mitochondrial proteins from various cell lines were electrophoresed through a BN- gel, electroblotted and hybridized with antibodies specific for subunits of complexes I, II and IV complexes (NDUFS2 antibody for complex I, SDHB antibody for complex II, and COX5A antibody for complex IV), and with TOM20 as a loading control. D Quantification of levels of complexes I, II, and IV. The calculations were based on three independent determinations in each cell line. Graph details and symbols are explained in the legend to Fig. 2