Mitochondrial Membrane Potential and Mass Assessment

A membrane potential probe, 3,3′-diethyloxacarbocyanine iodide (DiOC2(3)), (Thermo Fisher Scientific, Milan, Italy), was used to evaluate the mitochondrial membrane potential [37 (link),38 (link),39 (link)]. Cells were incubated with 10 μM DiOC2(3) for 30 min at 37 °C, washed twice, resuspended in PBS and analyzed by flow cytometry. To measure changes in the mitochondrial mass, cells were exposed to 200 nM MitoTracker Red CMXRos probe (Thermo Fisher Scientific, Milan, Italy) for 30 min at 37 °C, according to the manufacturer’s instructions. After two washes, labelled mitochondria were analyzed by flow cytometry.