Figure 7.

Effect of AST and ISO on the activity of complex II (CII) in rat heart mitochondria. (a) The upper part: freshly prepared rat mitochondria isolated from each group were solubilized in a buffer for BNE samples (Material and Methods section). Mitochondrial samples were separated by 1D BNE, and the gel was stained for the detection of the activity of CII by a buffer containing 50 mM KH₂PO₄ (pH 7.4), 84 mM succinate, 0.2 mM PMS (phenazine methosulfate), and 2 mg/mL NBT; the lower part: a diagram quantitatively reflecting changes in the activity of CIV; (b) the upper part: the excised bands with enzymatic activity were subjected to 2D SDS-PAGE and blotted onto nitrocellulose for immunodetection using the monoclonal antibody against SDHB (succinate dehydrogenase [ubiquinone] iron-sulfur subunit of CII); the lower part: a diagram quantitatively reflecting changes in the protein level. The data are presented as the means ± SDs of four independent experiments. * p < 0.05—significant difference in the protein level compared with the control (group 1), # p < 0.05 compared to RHM from ISO-injected rats (group 3). The statistical significance of the differences between the pairs of mean values was evaluated using the Student-Newman-Keul test (Supplementary Materials, p. 5).