Fig. 3: The effect of GLUT3 on GBM cells varies significantly in conditions with or without TMZ/CAPE treatment.

A GLUT3 KD efficiencies in U343-MG and LN-229 cell line. CCK8 (B) and colony formation C assays were performed to determine the cell proliferation ability of U87-MG cells, U343-MG cells, and LN-229 cells with or without GLUT3 KD and U251-MG cells with or without GLUT3 OE. D Transwell assay for invasion and migration was performed on U87-MG cells, U343-MG cells, and LN-229 cells with or without GLUT3 KD, and U251-MG cells with or without GLUT3 OE. E Wound healing assay was performed to detect the cell migration ability of U87-MG cells, U343-MG cells, and LN-229 cells with or without GLUT3 KD and U251-MG cells with or without GLUT3 OE. The wound closure percentage was calculated by Image J software. F 2-NBDG uptake by GLUT3 KD U87-MG, U343-MG, and LN-229 cells, as well as 2-NBDG uptake by GLUT3 OE U251-MG cells relative to control (Ctrl) cells. Cells were cultured with fluorescent probe 2-NBDG (100 μM) in glucose-free DMEM for 30 min at 37 °C; fluorescence signals were detected and normalized by cell number. G Colony formation assay was performed to determine cell proliferation ability. U87-MG, U343-MG, and LN-229 cells with or without GLUT3 KD were treated with DMSO or TMZ (200 μM) or CAPE (400 μM) once every two days until macroscopic clones were formed. Meanwhile, U251-MG cells with or without GLUT3 OE were manipulated as above. Clones were calculated by Image J software. Data are presented by means ± SEM of (n = 3) biologically independent experiments. One-way ANOVA with Tukey’s post-hoc test (A, B–F, U87-MG, U343-MG, LN-229 cell line, and G) or independent-sample t-tests (B–F, U251-MG cell line) 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).