Fig. 4.

Measurement of cellular in mitochondria. a Respiratory complex activities. The activities of respiratory complexes were investigated by enzymatic assay on complexes I, II, III, and IV in isolated mitochondria from lymphoblastoid cell lines derived from the mutant and control cybrid cell lines. Activities of complexes I, II, III, and IV were normalized by citrate synthase activity. b mitochondrial ATP levels. Mutant and control cell lines were incubated with 10 mM glucose or 5 mM 2-deoxy-d-glucose plus 5 mM pyruvate to determine ATP generation under mitochondrial ATP synthesis. Average rates of ATP level per cell line in mitochondria are show. The determinations were made for each cell line.The calculations were based on the independent determinations in each cell line. c Ratio of geometric mean intensity. Measurement of mitoROS. The levels of ROS generation by mitochondria in living cells from mutant and control cell lines were determined using the mitochondrial superoxide indicator MitoSOX-Red. The average of the determinations for each cell line is shown. The error bars indicate two standard errors of the means. p indicates the significance, according to the t-test, of the differences between mutant and control cell lines