Fig. 5
Overexpression of hENT1 in GEM-R cells and its reversal of chemoresistance in vitro. Note: Data are represented as mean ± SD. All experiments were performed in triplicate and repeated independently three times. A RT-PCR showed expression of hENT1 in mRNA level between WT and GEM-R cells. B RT-PCR showed overexpression of hENT1 in mRNA level in GEM-R-hENT1 cells compared with GEM-R cells. C Western blot showed expression of hENT1 in protein level of WT, GEM-R cells and GEM-R-hENT1 cells. Grayscale value ratio = The area of the target protein band/The area of the endogenous reference protein band. D CCK8 assay was used to evaluate IC50 of GEM-R cells and GEM-R- hENT1 cells to gradient concentration of GEM. E Percentage of cell apoptosis of GEM-R cells and GEM-R- hENT1 cells was evaluated by detection of flow cytometry with Annexin-V FITC/PI Apoptosis Detection Kit. F Uptake of GEM in WT, GEM-R and GEM-R- hENT1 cells by MRM analysis. Flow cytometric: Annexin V−/P− represented normal cells; Annexin V−/P + represented necrotic cells; Annexin V + /P + represented late apoptotic cells; Annexin V + /P- represented early apoptotic cells