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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Hypoxia potentiates gemcitabine-induced stemness in pancreatic cancer cells through AKT/Notch1 signaling

Fig. 6

AKT/Notch1 signaling mediates the synergistic enhancement of stemness induced by gemcitabine and hypoxia. (a) PANC-1 and Patu8988 cells were treated under hypoxic conditions (1%) for different durations, and the expression levels of NICD1, p-AKT, and AKT were determined by Western blot analysis. (b) After treatment with different concentrations of CoCl2, the changes in NICD1, p-AKT, and AKT expression levels were determined by Western blot analysis. (c) Two cell lines were co-treated with gemcitabine and CoCl2, and the protein expression levels were determined by Western blot analysis. (d) Two pancreatic cancer cell lines were pretreated with 10 μM DAPT or 20 μM LY294002 and then treated with a combination of gemcitabine and CoCl2. Then, the expression levels of Bmi1, Sox2, and NICD1 were determined by Western blot analysis. (e-g) After this treatment, the two cell lines were cultured with stem cell medium, and the ability of the cells for sphere formation was investigated by the sphere-forming assay: (e) Representative image of spheres after treatment; (f, g) Charts showing the data on sphere number and size. All data shown are from three independent experiments. Scale bar, 50 μm. *P < 0.05; **P < 0.01; ***P < 0.001

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