Fig. 3.

NFIB mRNA is a direct target of miR-346, and NFIB protein expression is inversely correlated with that of miR-346 in glioma tissues. a Left panel: predicted miR-346 target sequence in the wild-type (WT) 3′-UTR of NFIB mRNA and the mutated construct (mut). Right panel: Dual luciferase assay results. U87 and U251 cells were co-transfected with the recombinant luciferase plasmids. Luciferase activity of control group was normalized to 100%. b Spearman’s correlation analysis of NFIB protein and miR-346 expression levels in human glioma specimens (r² = 0.4442, P < 0.01). c NFIB mRNA expression level in miR-346 over-expressed cells, detected by real-time PCR. GAPDH was used as internal control. d Western blot analysis of NFIB protein expression in normal human astrocytes (NHAs) and U87, LN229, U251, A172, and U118 glioma cell lines. e Western blot analysis of NFIB protein expression levels in U87 and U251glioma cells transfected with miR-NC or miR-346. **P < 0.01, ***P < 0.001