Fig. 3

CBL0137-induced apoptosis and autophagy are dependent on mitochondrial ROS generation. A Jeko-1 cells treated with 1.0 μM or 2.0 μM CBL0137 were stained with the fluorescent mitochondrial probe JC-1, and the mitochondrial membrane potential (MMP) of the cells was observed by fluorescence microscope (40×, magnification). The quantitative results were analyzed. B The MMP stained with JC-1 probe was measured by flow cytometry. The histogram showed the percentage of JC-1 aggregate monomer. The data were expressed as mean ± SD (n = 3). C B-NHL cells were treated with different concentrations of CBL0137 for 24 h and then incubated with 10 μM DCFH-DA at 37 °C in the dark for 20 min, the fluorescent intensity was detected by flow cytometry. The mean fluorescence intensity of ROS was shown in histograms. The data were expressed as mean ± SD (n = 3). D B-NHL cells were preincubated with 2.5 mM NAC for 2 h and then treated with 2.0 μM CBL0137 for 24 h. The fluorescence intensity was detected by flow cytometry. E B-NHL cells were preincubated with 2.5 mM NAC for 2 h and then treated with 1.0 μM CBL0137 for 24 h. The changes in cell apoptosis rates were measured by flow cytometry. F In the presence or absence of NAC preincubation, the changes of apoptosis and autophagy-related proteins expression in B-NHL cells after CBL0137 treatment were detected by western blotting. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001