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Fig. 5 | Molecular Medicine

Fig. 5

From: Everolimus regulates the activity of gemcitabine-resistant pancreatic cancer cells by targeting the Warburg effect via PI3K/AKT/mTOR signaling

Fig. 5

Individual and combined effect of GEM and Evr on tube formation, migration, and apoptosis in GEMsen and GEMres cells. a Effect of GEM or/and Evr on tube formation in GEMres and GEMsen cells. Scale bar, 100 μm. b Transwell assay and quantification of the migration of GEMsen and GEMres cells treated with GEM or/and Evr after 48 h of culture. Scale bar, 100 μm. c Quantification of the number of migrating GEMsen and GEMres cells. The numbers in the arrows represent the difference (increase or decrease) in cell count between GEM and GEM + Evr. In GEM-treated GEMres cells, the enhancing effect of Evr on migration ability was stronger than that in GEM-treated GEMsen cells. d Ratio of the number of migrated GEMres cells to that of migrated GEMsen cells. e Flow cytometry of apoptosis in GEMsen and GEMres cells treated with GEM or/and Evr. Numbers in the upper right quadrant denote the percentage of cells in late apoptosis. f Quantification of the proportion of late-apoptotic cells. The numbers in the arrows represent the difference (increase or decrease) in the percentage of late apoptosis between GEM and GEM + Evr. In GEM-treated GEMres cells, the enhancing effect of Evr on apoptosis was stronger than that in GEM-treated GEMsen cells. g Ratio of the apoptosis of GEMres cells to that of GEMsen cells. h Western blot of proteins associated with apoptosis. Quantification of the protein expression of i Bax, j Bcl-2, and k Cyt-c in GEMsen and GEMres cells treated with GEM or/and Evr, normalized to that of GAPDH as an internal control. From i to k, the numbers in the arrows represent the difference (increase or decrease) in protein expression between GEM and GEM + Evr. Compared to GEM-treated GEMsen cells, the effect of Evr on Bax and Bcl-2 expression was weaker in GEM-treated GEMres cells, but Evr exerted similar effect on Cyt-c expression. The data are expressed as the mean ± standard deviation of three replicates (n = 3). *P < 0.05; #P < 0.05 compared with the same treatment in GEMsen cells. GEM: gemcitabine, Evr: everolimus; GEMsen: GEM-sensitive pancreatic cancer cells; GEMres: GEM-resistant pancreatic cancer cells; au: arbitrary units

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