Figure 2. Flow cytometry analysis of mitochondrial function.

(A) Measurement of mitochondrial membrane potential. Cells were washed, resuspended in PBS containing 1% BSA with 20 nM DiOC₆ and 5 μg/ml PI and incubated for 15 min at 37 °C. A minimum of 20000 events were analysed for each sample as described in the Materials and methods section. (B) Measurement of ROS levels. Cells were washed, harvested and resuspended in PBS containing 1% BSA and 5 μM CMH₂DCFDA for 20 min at 37 °C. A minimum of 20000 events were analysed for each sample as described in the Materials and methods section. Results shown are representative of 2–3 separate experiments. Clear histograms show control cybrids and black histograms show mutant cybrids.