Figure 1

The effects of pharmacological inhibition of complexes I and II on the respirasome assembly, mitochondrial swelling and ROS production in cardiac mitochondria. (A,B), Representative images of BN-PAGE analysis of respirasomes in the presence of rotenone (A) and malonate (B) at given concentrations. (C,D), Quantitative data of respirasome levels. Respirasome levels were calculated as integrated pixel densities from the bands containing complex I, III, and IV (marked as vertical double headed arrows in (A,B) which were normalized by whole lane density. Bands were identified as shown previously^13,14. Data were normalized to control (non-treated) mitochondria. E. Mitochondrial swelling, as a marker PTP opening, measured by the decrease in absorbance at 540 nm. External Ca²⁺ (100 µM per arrow) was added to initiate swelling of mitochondria with or without 0.5 µM rotenone or 2.5 mM malonate. F. The rate of ROS production measured by Amplex Red and shown as pmol H₂O₂ per min to mg of mitochondrial protein. Results are presented as mean ± SE. *P < 0.05 vs. control. n = 3 per group.