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Fig. 1 | Cancer Cell International

Fig. 1

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

Fig. 1

Gambogic acid (GA) induces autophagy in pancreatic cancer cells. a PANC-1 and BxPC-3 cells were treated with the indicated concentrations of GA for 24 h, or with 1 µM GA for the indicated times. The expression of LC3 and P62 proteins was analyzed by western blot. b PANC-1 cells were treated with 1 µM GA for 24 h, and transmission electron microscopy was used to visualize autophagic vacuoles. c PANC-1 and BxPC-3 cells were treated with 1 µM GA in either the absence or presence of bafilomycin A1 (Baf-A1) (100 nM) for 24 h. Acidic vesicular organelles (AVOs) were detected with acridine staining, and the number of cells with AVOs was quantified. Scale bars indicate 200 µm. d Stable GFP-mRFP-LC3 transfected PANC-1 cells were treated with 1 µM GA in the absence or presence of 3-methyladenine (3-MA) (10 mM) for 24 h, and cells were observed under a fluorescence microscope. The number of autophagosomes (yellow dots) and autolysosomes (red dots) within each cell were counted; n = 50 cells/sample. e PANC-1 and BxPC-3 cells were treated with 1 µM GA in the absence or presence of 3-MA (10 mM), or chloroquine (CQ) (40 µM) for 24 h. The LC3 protein was analyzed with western blot. Data are presented as mean ± SD (n = 3); *** indicates P < 0.001

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