Fig. 5.

Inhibition of autophagy enhanced apoptosis induced by ethanol extracts of Chrysanthemum zawadskii Herbich (ECZ) by increasing the production of reactive oxygen species (ROS) in mouse colon cancer CT-26 cells. a Treatment with 3-methyladenine (3-MA) enhanced ECZ-induced apoptosis. b Effects of 3-MA treatment on caspase-3 and poly (ADP-ribose) polymerase (PARP) expression. c Relative fold change of cleaved caspase-3. d Relative fold change of PARP. Cells were pretreated with 5 mM 3-MA prior to 1 h. Apoptotic cells were evaluated by flow cytometry, and the proteins were detected by Western blot analysis. e Effects of 3-MA on ROS production. Cells were treated with 300 μg/ml ECZ for 24 h prior to 1 h incubation with 5 mM 3-MA or/and 5 mM NAC. ROS production was evaluated by flow cytometry. Data are presented as the mean ± standard deviation for at least three independent experiments. ***p < 0.001 vs. the control group, #p < 0.05, ^##p < 0.01 vs. the ECZ treated group