Fig. 4

miR-429 enhances TGF-β-induced EMT and invasion in NSCLC cells. a After being serum-starved for 24 h, A549 and H226 cells transiently overexpressing miR-429 were treated with or without TGF-β1 (5 ng/ml) for 1 h and 2 h, respectively. Snail mRNA expression was quantified by qRT-PCR analysis. Snail mRNA level of the unstimulated cells was assigned the value 1, and the relative Snail mRNA expression in TGF-β1-stimulated cells was recalculated accordingly. b After being serum-starved for 24 h, A549 and H226 cells transiently overexpressing miR-429 were treated with or without TGF-β1 (5 ng/ml) for 24 h and 48 h, respectively. Western blot analysis was performed to examine the expression of N-cadherin, which was normalized to β-actin. c A549 and H226 cells transiently overexpressing miR-429 were treated as above and allowed to migrate through an 8-μM pore in transwells. Migrated cells were stained and counted in at least three light microscopic fields. Scale bar, 100 μm. d Cells were treated as above and allowed to invade through Matrigel-coated membrane in transwells. Invasive cells were stained and counted under a light microscope. Scale bar, 100 μm. e After being serum-starved for 24 h, A549 and H226 cells transiently overexpressing anti-miR-429 were treated with or without TGF-β1 (5 ng/ml) for 1 h and 2 h, respectively. qRT-PCR analysis was done to determine the relative Snail mRNA expression. f After being serum-starved for 24 h, A549 and H226 cells transiently overexpressing anti-miR-429 were treated with or without TGF-β1 (5 ng/ml) for 24 h and 48 h, respectively. N-cadherin expression was analyzed by western blot. g A549 and H226 cells transiently overexpressing anti-miR-429 were treated as above and allowed to migrate through an 8-μM pore in transwells. Migrated cells were stained and counted in at least three light microscopic fields. Scale bar, 100 μm. h Cells were treated as above and allowed to invade through Matrigel-coated membrane in transwells. Invasive cells were stained and counted under a light microscope. Scale bar, 100 μm. **P < 0.01; ***P < 0.001