Bmi-1 promotes an aggressive phenotype in glioma cells via activation of the NF-kappaB-MMP-9 pathway. Bmi-1-overexpressing cells were treated with a NF-kappaB inhibitor JSH-23 at 30μM or MMP-9 inhibitor at 50μM. A, Quantification of cell migration in the Transwell assay (without matrigel). B, Quantification of cell invasion in the Transwell matrix penetration assay (with matrigel). C, Representative micrographs of the 3-D spheroid invasion assay. The experiments were repeated for at least three times with similar results. D, ELISA analysis of MMP-9 secretion. Error bars represent SD of three independent experiments; ** P < 0.01.