Figure 4
Alu RNA acts as a molecular decoy for miR-566 inducing EMT. (a) Alu RNA transfection in SW480 cells reduces miR-566 level as evaluated by quantitative reverse transcriptase (qRT–PCR). (b) SW620 cells show less miR-566 level compared with SW480 cells, as evaluated by qRT–PCR. (c) The activity of Renilla luciferase (RLuc) was downregulated by miR-566 and not by NT miRNA. SW480 cells were co-transfected with psiCHECK2 and/or psiCHECK2/Alu, and miR-566 and/or NT miRNA. The bars indicate Firefly luciferase (FLuc) activities normalized against Renilla. Each experiment was repeated at least three times and each sample was assayed in triplicate. (d) miR-566 prevents the Alu-induced EMT, as shown by Fibronectin, E-cadherin, TWIST1 and ZEB1 levels evaluated by western blot. Densitometric values normalized against β-Tubulin are shown in parentheses. (e) Representative images of F-actin (red) and E-cadherin (green) staining. Nuclei are counterstained with 4',6-diamidino-2-phenylindole (DAPI, blue). Scale bar: 75 μm.
