Fig. 4

AKT promotes pancreatic cancer cell stemness partly by mediating Notch1 activation. Two cell lines were pretreated with 20 μM LY294002 for 2 h and then treated with gemcitabine. (a) The expression levels of Bmi1, Sox2, NICD1, p-AKT (serine 473), and AKT were determined by Western blot analysis. (b) The change in the proportion of CD24⁺ pancreatic CSCs was determined by FCM. (c-e) The ability of the cells for sphere formation was investigated by the sphere-forming assay: (c) Representative image of spheres formed after treatment; (d and e) Charts showing the data on sphere number and size. The graphs show the results of three independent experiments. Scale bar, 50 μm. *P < 0.05; **P < 0.01; ***P < 0.001