Fig. 6

HDAC2 knockdown inhibits tumor-sphere formation and proliferation via increasing miR-3189-mediate GLUT3 in GSCs. A Glucose uptake in DOX-inducible shHDAC2 GSCs. B PARP cleavage in DOX-inducible shHDAC2 GSCs w/wo doxycycline was analyzed using western blot with indicated antibodies. C Tumor-sphere formation assay in GSCs. Cell picture images were taken at 40X. Scale bar: 400 μm. D Quantification of tumor-sphere number. E Cell viability in miR-3189-transfected GSCs by WST-8 assays. F mRNA expression of GLUT3 in miR-3189-expressing GSCs using qPCR. G Luciferase reporter activity in DOX-inducible shHDAC2 GSCs using dual-luciferase assay. GSCs were transiently transfected with pmirGLO-GLUT3MT-Luc. Reporter activities were normalized relative to Renilla luciferase activities. Reporter activities were expressed as the mean ± SD for triplicates. H Cell proliferation of shHDAC2 expressing GSCs. I Graphical conclusion of the mechanism of cell death in HDAC2 knockdown GBM/GSCs. All data are expressed as the mean ± SD from three independent experiments, each performed in triplicate. *p < 0.05. **p < 0.01, ***p < 0.001