Figure 5 | Scientific Reports

Figure 5

From: Mechanistic Evaluation and Translational Signature of Gemcitabine-induced Chemoresistance by Quantitative Phosphoproteomics Analysis with iTRAQ Labeling Mass Spectrometry

Figure 5

Effects of targeted anticancer agents on Patu8988 cells. (a) Patu8988 cells were treated with gemcitabine for 48 h at the concentrations of 2.5 μg/mL, and the expression of phosphorylated proteins were detected by Western Blot. (b,c) The pancreatic CSCs markers CD133-PE, CD24-FITC after gemcitabine and the corresponding inhibitors/activator treatment were measured by flow cytometry. The cells were treatment with gemcitabine (2.5 μg/mL, 48 h), rapamycin (1 μM, 48 h), LY294002 (20 μM, 24 h) and A-769662(20 μM, 2 h). The numerical values represent the percentage of positive cells. (d) After inhibitors or activator treatment, the Patu8988 cells were performed with the tumor sphere-forming experiments in stem cell medium. (e,f) Dot-plots represented the apoptotic status of Patu8988 cells using Annexin V-FITC/PI method. The dot-plots in the upper right and lower right quadrant were considered as the percentage of apoptotic cells. (g) The expression levels of Bmi1, Nanog and Sox2 were compared with western blot analysis after gemcitabine and the corresponding inhibitors/activator combination treatment. The results were representative of three independent experiments. Scale bar, 200 μm. *P < 0.05; **P < 0.01.

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