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Determination of Mitochondrial Reactive Oxygen Species (ROS)
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HepG2 cells were treated for 24 h to quantify mitochondrial ROS according to the specified protocol. After treatment, cells were incubated with MitoSOX Red and MitoTracker Green reagents at 37 °C for 15 min. Subsequently, the cells were washed with PBS to remove the excess probe. Fluorescence microscopy (Nikon, Tokyo, Japan) was used to analyze the cells. The intensity of MitoSOX Red fluorescence was directly correlated with the level of mitochondrial ROS production.
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