Figure 4: Inhibition of miR-497-5p suppresses TGF-β1-induced myofibroblast differentiation of lung resident mesenchymal stem cells (LR-MSCs).

(A) LR-MSCs were transfected with either 5 × 10⁷ TU/ml negative control inhibitor (LV-NC-inhibitor) or miR-497-5p inhibitor (LV-miR-497-5p-inhibitor), followed with treatment either with or without 10 ng/ml TGF-β1 for 7 days. The levels of miR-497-5p, Reck, Acta2 and Col1a1 were measured by quantitative real-rime polymerase chain reaction. Three independent experiments were performed, and the values are expressed as the means ± SD. *P < 0.05 and **P < 0.01. (B) The experiments were performed as in (A). The expression of Reck, Mmp2, Mmp9, Tgfb1, Fn1, Col1a1, Vim and Acta2 was determined by Western blot. The expression levels of protein were quantified by densitometry and normalized to the expression of β-actin. Three repeats were performed. *P < 0.05 and **P < 0.01. (C) LR-MSCs were treated as explained in (Figs 3A and 4A), the expression of Acta2 and Col1a1 was measured by immunofluorescence. Acta2 and Col1a1were revealed with secondary 594-labeled antibodies; nuclei were revealed by DAPI staining.