Fig 1: Western blot analysis of mitochondrial proteins. Twenty micrograms of total cellular proteins from various cell lines was electrophoresed through a denaturing polyacrylamide gel, electroblotted and hybridized with antibodies for (A) 8 mtDNA-encoding (ND1, ND3, ND4, ND4L, ND5, CYTB, CO2 and ATP8) and (B) 10 nucleus-encoding (NDUFA1, NDUFS1, NDUFA10, SDHB, UQCRC2, UQCRFS1, COXIV, COX17, ATP5B and ATP5F1) subunits of OXPHOS (CI, CII, CIII, CIV and CV represented complexes I, II, III, IV and V, respetively), TOM20 or GAPDH as a loading control, respectively. (C) Quantification of mitochondrial protein levels. Average relative ND1, ND3, ND4, ND4L, ND5, CYTB, CO2, and ATP8 content per cell, normalized to the average content per cell of TOM20 in TRUB1KO and WT cell lines. The values for the TRUB1KO cell lines are expressed as percentages of the values for the WT cell line. The error bars indicate two standard errors of the means, the horizontal dashed lines represent the average value for each group. The calculations were based on three independent determinations in each cell line. P indicates the significance, according to the t-test, of the differences between TRUB1KO and WT cell lines.