Figure 3
Antisense oligonucleotide (Alu AS) targeting Alu RNA sequences rescues the DICER1 deficit-induced EMT. (a) Alu AS reduces Alu RNA accumulation induced by DICER1 deficit in SW480 cells as evaluated by quantitative reverse transcriptase (qRT–PCR). (b) Evaluation of DICER1, Vimentin, Fibronectin, E-cadherin, TWIST1 and ZEB1 protein levels by western blot analysis. Densitometric values normalized against β-Tubulin are shown in parentheses. (c) Representative images of F-actin (red), E-cadherin (green) and Vimentin (red) staining. Nuclei are counterstained with 4',6-diamidino-2-phenylindole (DAPI, blue). Scale bar: 50 μm. (d and e) Alu AS inhibits DICER1-deficit induced SW480 cell migration and invasion. Data are expressed as fold of induction with respect to Ctrl AS. (f) DICER1 AS induced global miRNA expression deficits in SW480 cells compared with Ctrl AS. No significant difference in miRNA abundance between Alu AS and Ctrl AS-treated DICER1-depleted cells as evaluated by qRT–PCR. For all panels: n=3; *P<0.05. Error bars denote s.e.m.
