Fig. 4

Reactive oxygen species (ROS) regulated apoptosis and autophagy induced by ethanol extracts of Chrysanthemum zawadskii Herbich (ECZ) in mouse colon cancer CT-26 cells. a Representative histogram of intracellular ROS levels. Cells were treated with various concentrations (200–300 μg/ml) of ECZ for 24 h, and dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence intensity was detected by flow cytometry. b N-Acetyl-L-cysteine (NAC) attenuated the increased ROS production induced by ECZ. (C) NAC attenuated apoptosis induced by ECZ. d Effects of NAC on procaspase-3 and poly (ADP-ribose) polymerase (PARP) expression. e Relative fold change of cleaved caspase-3. f Relative fold change of cleaved PARP. g Effects of NAC on autophagy. h Effects of NAC on LC3B and p62/SQSTM1 expression. i Relative fold change of LC3B-II. j Relative fold change of p62/SQSTM1. Cells were treated with 300 μg/ml of ECZ for 24 h prior to 1 h incubation with 1 mM NAC. ROS production, apoptotic cells, and autophagic cells were detected by flow cytometry. Total lysates were subjected to SDS-PAGE for Western blot analysis. 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, ^###p < 0.001 vs. the ECZ treated group