Research Paper Volume 13, Issue 4 pp 5055—5068

Figure 3. miR-29a-3p inhibited migration and VM formation by directly targeting ROBO1. (A) Schematic representation of the predicted binding sites for miR-29a-3p in the ROBO1 3′-UTR (wild type; WT) and the designed mutant versions (mutant; MUT) of the ROBO1 3’-UTR (left panel). Relative luciferase activity of HEK293T cells in the presence of the indicated treatments (middle and right plots). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05). (B) Western blot analysis of the protein level of ROBO1,N-cadherin, MMP9 and LAMB2 after miR-29a-3p transfection. Results are from three independent experiments. (C) Western blot analysis of the expression of ROBO1,N-cadherin, MMP9 and LAMB2 after ROBO1 and miR-29a-3p overexpression. Results are from three independent experiments. (D) Representative images for the transwell assay (scale bar, 100 μm; n=3). (E) Quantification of transwell migration assays in (D). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05). (F) Representative images for the VM formation assay (scale bar, 200 μm; n=3). (G) Quantification of relative VM numbers in (F). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05).