Fig. 6

miR-29a activates a Sox4/HIC5 invasion pathway in glioblastoma. a) Effect of lentiviral-mediated overexpression of miR-29a on Sox4 expression in LN229, U87, U251 or primary human glioblastoma stem-like cells (GSC). b) Effect of miR-29a antagomiR (100 nM) on Sox4 protein expression in primary human glioblastoma stem-like cells. c) Western blot illustrating Sox4 knockdown using siRNA transfection in LN229 or U251 glioblastoma cells. d) Phase micrographs (left panels) or quantitation of data (right panel) from a Matrigel invasion assay showing effect of siRNA-mediated Sox4 knockdown on LN229 or U251 glioblastoma cell invasion. Mean ± SEM. P < 0.0001, unpaired t-test. Scale approximately 50 μm. e) Phase micrographs (left panels) or data quantitation (right panel) from a 3D Matrigel invasion assay showing effect of Sox4 overexpression on PTEN-deficient U87 glioblastoma cell invasion. Mean ± SEM; P < 0.01, unpaired t-test. Scale approximately 50 μm. f) Effect of miR-29a mimic or Sox4 siRNA on HIC5 protein expression in human U251 glioblastoma cells. g) Phase micrographs (left panels) or data quantitation (right panel) from a Matrigel invasion assay showing effect of siRNA-mediated HIC5 knockdown on LN229 or U251 glioblastoma cell invasion. Mean ± SEM. *P < 0.0001, unpaired t-test. Scale approximately 50 μm. h) Phase micrographs (left panels) or data quantitation (right panel) from a Matrigel invasion assay showing effect of siRNA-mediated HIC5 knockdown on invasion induced by miR-29a in human U251 glioblastoma cells. Mean ± SEM. *P < 0.0001, unpaired t-test. Scale approximately 50 μm