FIG. 5.

The role of ROS in JDA-202-induced esophageal cancer cell death. Four esophageal cells were treated with JDA-202 for 12 h. The levels of ROS and H₂O₂ (A, B) were detected by flow cytometry. After pretreatment with or without 5 mM NAC or 1 or 2 U/L catalase for 2 h, EC109 cells were treated with indicated concentrations of JDA-202 for 12 h; then, the intracellular ROS was detected by a fluorescence microscope (C, F) or flow cytometry (D, E, G, H). (I) After pretreatment with or without 5 mM NAC or 2 U/L catalase for 2 h, EC109 cells were treated with indicated concentrations of JDA-202 for 24 h; cell viability was determined by MTT assay, and ΔΨm was detected by flow cytometry (J) and quantitated (K). (L) After pretreatment with or without 1 or 2 U/L catalase for 2 h, EC109 cells were treated with indicated concentrations of JDA-202 for 24 h, and ΔΨm was detected by a fluorescence microscope. After pretreatment with or without 5 mM NAC for 2 h, EC109 cells were treated with indicated concentrations of JDA-202 for 24 h, and the expression and quantitation of Bcl-2 and Bax (M, N) were detected by Western blot and Image J. Data are presented as means ± SD. Three individual experiments were performed for each group. ^**,##,$$p < 0.01, compared with the controls. ΔΨm, mitochondrial membrane potential; NAC, N-acetylcysteine; ROS, reactive oxygen species. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars