Figure 5.

Ursolic acid induced autophagy by reactive oxygen species (ROS) induction in ESCC. (A) The UA-induced ROS-mediated DCFDA fluorescence intensity of the cells treated with UA (30 µM) increased in a time-dependent manner. (B) The quantitative analysis of ROS by fluorescence-activated cell sorting showed increased DCFDA fluorescence in a dose-dependent manner. (C) DPI- and NAC-treated cells showed decreased fluorescence intensity, demonstrating intracellular ROS inhibition. (D) Combination treatment with the ROS inhibitor DPI increased cell viability compared with UA treatment alone. (E) TE-8 and TE-12 cells were treated with 30 µM UA and 100 nM or 250 nM DPI. We detected LC3 protein levels by Western blot analysis and employed GAPDH as an internal control. The data are presented as the mean ± SE for three independent experiments.*** p < 0.001 compared with the control, ^## p < 0.01, and ^### p < 0.001 compare with the UA-treated group.