Figure 9.

Effect of AST and ISO on the activity of ATP synthase in rat heart mitochondria. (a) The upper part: freshly prepared rat mitochondria isolated from each group were solubilized in the buffer for BNE samples (Materials and Methods section). Mitochondrial samples were separated by 1D BNE, and the gel was stained for 16 h for the detection of complex V (CV) activity by a buffer containing 10 mM ATP, 35 mM Tris (HCL), 270 mM glycine, 14 mM MgSO₄, and 0.2% Pb(NO₃)₂; the lower part: a diagram quantitatively reflecting changes in CV activity; (b) the upper part: excised bands with enzymatic activity were subjected to 2D SDS-PAGE and blotted onto nitrocellulose for immunodetection using monoclonal antibodies against subunits α (ATP5A), c (ATP5G), b (ATP5F1), and CyP-D; the lower part: a diagram quantitatively reflecting changes in the protein levels. 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. 7).