Figure 5
TGF-β1 stimulation induces Alu RNA accumulation. (a) The stimulation of SW480 cells with 5 ng/ml TGF-β1 induces EMT. Representative images (upper) of TGF-β1 (right) or not- (NT, left) treated SW480 cells stained for F-actin (red) and E-cadherin (green). Nuclei are counterstained with 4',6-diamidino-2-phenylindole (DAPI, blue). Scale bar: 75 μm. Western blot (lower) shows abundance of Fibronectin, E-cadherin, TWIST1 and ZEB1. Densitometric values normalized against β-Tubulin are shown in parentheses. (b) TGF-β1 increases Alu RNA accumulation as evaluated by northern blot (upper) and quantitative reverse transcriptase (qRT–PCR) (lower). (c) TGF-β1 decreases miR-566 in SW480 cells, as evaluated by qRT–PCR. (d and e) miR-566 and Alu AS rescues the TGF-β1-induced EMT. Western blots show abundance of Fibronectin, E-cadherin, TWIST1 and ZEB1. Densitometric values normalized against β-Tubulin are shown in parentheses. For all panels: n=3; *P<0.05. Error bars denote s.e.m.
