Combined effects of CGA, TC-HT, and LIPEF on ROS generation in PANC-1 cells.
(A) The DHE (O2•−) levels were measured by flow cytometry after treatment with CGA (200 μM), TC-HT (10-cycles), and LIPEF (60 V/cm) alone, in double or triple combination on PANC-1 cells for 24 h. (B) Graph shows the fold change in MFI of DHE relative to control. (C) The PANC-1 cells were treated with the same parameters of CGA, TC-HT, and LIPEF alone or in combination with or without NAC (5 mM). The cell viability was measured by MTT assay and shown as a percentage of the control. (D) The expression levels of p53, p21, Bax, and Bcl-2 were examined by western blot analysis after the triple treatment with or without NAC (5 mM). GAPDH was used as internal control. The expression levels of these proteins with or without the 5 mM NAC pretreatment were measured and shown in (E), where the expression data were normalized to GAPDH for comparison. Data are presented as mean ± S.D. in triplicate. (***p < 0.001 vs. untreated control; #p < 0.05, ##p <0.01, and ###p < 0.001 between indicated groups).