2.5. Reactive Oxygen Species (ROS) Generation Assay

The intracellular ROS levels were evaluated using the ROS-specific fluorescent dye H2DCF-DA. After indicating treatments, cells were washed with PBS for three times and stained with H2DCF-DA, incubated at 37°C for 30 min in the darkness. Furthermore, cells were harvested and the generation of ROS was quantified by measuring the intracellular fluorescence intensity by a flow cytometer. The samples were then analyzed with FlowJo 7.6.1.