Measurement of mitochondrial ROS by flow cytometry

Mitochondrial ROS generation was determined by flow cytometry with MitoSOX Red Mitochondrial Superoxide Indicator. The samples were collected and stained with 5 mircoM MitoSOX and then were incubated in the dark in a water bath at 37°C for 15 min. The cell suspension was then transferred to a 5 ml FACS tube and analyzed on a flow cytometer within 10 min using Cell Quest software (BD Biosciences, Franklin Lakes, NJ, USA).