Fig. 2

Signaling pathway associated with enhanced migration in miR-200b/200a/429 or miR-141/200c-transduced MDA-MB-231 cells. a, b Western blotting analysis of the levels of phosphorylated FAK, AKT, ERK, and integrin-αV expression. The levels of FAK and AKT phosphorylation were significantly higher in the miR-200 family-transduced cells than in the control cells, but the level of ERK phosphorylation was similar between the miR-200 family-transduced cells and control cells. The miR-200 family-transduction also increased integrin-αV expression. Densitometric quantifications of FAK, AKT, and ERK phosphorylation in the miR-200 family-transduced cells relative to control cells. β − actin was used as an internal reference. c Immunofluorescence analysis of phosphorylated FAK and integrin-αV. Phosphorylated FAK and integrin-αV were overexpressed and were highly localized at membrane surfaces in the miR-200 family-transduced cells. d Trans-well migration assay of FAK (PF573228) and PI3K/AKT (LY294002)-inhibitor-treated cells. Treatment with inhibitors of the FAK and PI3K/AKT signaling pathways significantly suppressed the cell migration enhanced by the transduction of miR-200 family. All experiments were performed at least in triplicate, and the values are the mean values ± standard deviation. *p < 0.05 **p < 0.001, Scale bar, 10 μm