Figure 3.

PTBP3 mediates TGF-β-induced EMT, migration and invasion of LUAD cell lines. (a) A549 cells were transfected with scramble and PTBP3 siRNA (100 nM) for 48 h and then treated with TGF-β1 (5 ng/ml) for 24 h, subsequently, the cells underwent immunofluorescence analysis by using E-cadherin and Vimentin antibodies (red). Cell nuclei were stained with DAPI (blue). scale bars, 20 μm. (b) A549 and H1299 cells transfected with scramble and PTBP3 siRNA were treated with 5 ng/ml or 10 ng/ml TGF-β1 for 24 h. Then EMT markers E-cadherin, N-cadherin and Vimentin expression were analyzed using Western blot. GAPDH was used for internal control. (c-f) A549 and H1299 cells were transfected as above. Whole cell lysates were analyzed by Western blot (c and e) and qRT-PCR (d and f) for the indicated genes. qRT-PCR data were normalized to GAPDH and presented as mean ± SD from three independent experiments. ***P < 0.001 by ANOVA. (g and h) PTBP3-knowdown A549 and H1299 cells and control cells were treated with or without TGF-β1, and were subjected to the transwell migration and invasion assay. Data were presented as the mean ±SD of triplicate experiments. *P < 0.05, **P < 0.01 by Student’s t-test.