Fig. 4

GLUT3 knockdown by miR-3189 increased cell death of GBM cells. A FACS analysis of GBM cells expressing control siRNA, HDAC2 siRNA, GLUT3 siRNA, control miRNA, and miR-3189 mimics. B TUNEL assay in HDAC2 siRNA, GLUT3 siRNA, control miRNA, and miR-3189 mimics-transfected GBM cells. TUNEL-positive cells: green, Nuclei with PI: red, Merged cells: yellow. Scale bar: 100 μm. C Colony formation assay of miR-3189 mimic-transfected GBM cells. Quantification of colony formation. D Schematic of miR-3189 and GLUT3 3′-UTR binding site interactions. E Measurement of luciferase reporter activity of pmirGLO-GLUT3-Luc for binding of miR-3189. pmirGLO-GLUT3MT-Luc plasmid were transiently transfected w/wo miR-3189 mimics into 293 T and GBM cells. Reporter activities were normalized relative to Renilla luciferase activities. F Luciferase reporter assays using pmirGLO-GLUT3MT-Luc plasmid transiently transfected into DOX-inducible shHDAC2 GBM cells. Reporter activities were normalized relative to Renilla luciferase activities. All data are expressed as the mean ± SD from three independent experiments, each performed in triplicate. **p < 0.01, ***p < 0.001