Fig. 6: GLUT3 is a TMZ-CAPE-glucose multifunctional transporter.
From: GLUT3 enhances chemosensitivity in glioblastoma by transporting temozolomide and capecitabine
A, B Binding of GLUT3 with glucose (upper panels), TMZ (middle panels), or CAPE (lower panels) detected by MST assay. Cell lysates used in the MST assay were obtained from U251-MG (A) and U343-MG (B) cells stably expressing GLUT3-GFP and corresponding control, and the assays were performed at pH 6.8 or 7.4, respectively. Binding curves and Kd (dissociation constant) values are also shown. C GFP-GLUT3 OE efficiency in U251-MG (upper panel) and U343-MG (lower panel) cell lines. D Detection of TMZ binding to GLUT3 WT or its mutants by MST assay. E Comparison of TMZ uptake by U251-MG cells stably expressing GLUT3 mutants and WT GLUT3. F Detection of CAPE binding to GLUT3-GFP WT or its mutants by MST assay. G Comparison of CAPE uptake by U251-MG cells stably expressing GLUT3 mutants and WT GLUT3. Effect of GLUT3 mutations on IC50 values of TMZ (H) and CAPE (I) in U251-MG cells detected by CCK-8 assay. J Effect of GLUT3 mutations on colony formation of U251-MG cells. Cells overexpressed with GLUT3 Q159A, W386A, or WT were treated with DMSO, TMZ (200 μM), or CAPE (400 μM) once every two days until macroscopic clones were formed. Clones were calculated by Image J software. Data are presented by means ± SEM of (n = 3) (A–D, F, H–J) or (n = 6) E, G biologically independent experiments. Independent-sample t-tests (C, H–J) or one-way ANOVA with Tukey’s post-hoc test E, G were used for statistical analysis by using SPSS 20 (IBM). A p-value < 0.05 was considered significant (*P < 0.05, **P < 0.01, ***P < 0.001).
