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Fig. 6 | Cancer Cell International

Fig. 6

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

Fig. 6

Inhibition of autophagy enhanced ROS production in pancreatic cancer cells treated with gambogic acid (GA). a PANC-1 and BxPC-3 cells were pretreated with 40 µM chloroquine (CQ) for 24 h, and then treated with 2 µM GA for another 24 h in the absence or presence of N-acetylcysteine (NAC) (20 mM), and the mitochondrial membrane potential was measured using the JC-1 assay. bd PANC-1 and BxPC-3 cells were pretreated with 40 µM CQ for 24 h, and then treated with 1 µM GA for another 24 h in the absence or presence of NAC (20 mM). Reactive oxygen species (ROS) production was detected by CM-H2DCFDA staining and observed under a fluorescence microscope (×100) (b). ROS production was detected by CM-H2DCFDA staining and ROS levels were measured using a fluorescence microplate (c). Cell viability was detected by the MTT assay (d). e PANC-1 and BxPC-3 cells were pretreated with 40 µM CQ for 24 h, and then treated with 2 µM GA for another 24 h in the absence or presence of NAC (20 mM), and the expression of cleaved caspase-9, cleaved-PARP, cleaved caspase-3, and LC3 proteins was analyzed with western blot. Data are presented as mean ± SD (n = 3); *** indicates P < 0.001

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