Reactive Oxygen Species Measurements

The activity of ROS was detected using the DCFH-DA kit (S0033, Beyotime) following the manufacturer's guidebook. 2′,7′-Dichlorodihydrofluorescein diacetate was used as the oxidation-sensitive fluorescent probe. PC cells (1-5 × 10⁶/ml) were incubated with a probe at 37°C for 20 min in a 35 mm porous glass bottom dish (Cellvis, USA) and the nuclei were stained using Hoechst (Beyotime). A laser scanning confocal microscope (Nikon, Japan) was used to capture the image. Quantitative DCF fluorescence was measured by taking the average of measurements obtained on multiple views.