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Fig. 5 | Cancer Cell International

Fig. 5

From: Gambogic acid induces autophagy and combines synergistically with chloroquine to suppress pancreatic cancer by increasing the accumulation of reactive oxygen species

Fig. 5

Gambogic acid (GA) promotes ROS generation by damaging mitochondria, and induces pancreatic cancer cell autophagy. a PANC-1 and BxPC-3 cells were either treated with the indicated concentrations of GA for 24 h, or treated with GA for the indicated times. The generation of reactive oxygen species (ROS) was detected by CM-H2DCFDA staining and observed under a fluorescence microscopes (×100). b ROS production was detected by CM-H2DCFDA staining and ROS levels were measured using a fluorescence microplate. c PANC-1 and BxPC-3 cells were treated with 1 µM or 2 µM GA for 24 h, and the mitochondrial membrane potential was measured using the JC-1 assay. d PANC-1 and BxPC-3 cells were treated with 1 µM GA in the absence or presence of N-acetylcysteine (NAC) (20 mM) for 24 h. The acidic vesicular organelles (AVOs) were detected with acridine staining and observed under a fluorescence microscope. Scale bars indicate 200 µm. e PANC-1 and BxPC-3 cells were treated with 2 µM GA in the absence or presence of NAC (20 mM) for 24 h, and the expression of cleaved caspase-9, cleaved-PARP, and LC3 proteins was analyzed with western blot. f PANC-1 and BxPC-3 cells were treated with GA (2 µM) for 24 h in the presence or absence of NAC (20 nM). Apoptosis was detected using the Annexin V/PI double stain, and flow cytometry was performed. Data are presented as mean ± SD (n = 3); *** indicates P < 0.001

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