SMAD3 mediates PDLIM5 function in cancer cells.
A, Western blotting analysis of PAI1 and JUNB in PDLIM5 knockdown A549 cells, with or without SMAD3 overexpression. PAI1 and JUNB were quantified and normalized to shScr value (n = 3). β-Actin was used as a loading control. B, RT-PCR analysis of EMT markers in PDLIM5 knockdown A549 cells, with or without SMAD3 overexpression. The cells were treated, or not, with TGFβ1 (5 ng/ml) for 24 h. The data were normalized to 18S RNA (n = 3). C, representative images of the Transwell migration and Transwell invasion assay of PDLIM5 knockdown A549 cells with or without SMAD3 overexpression. Scale bar, 200 μm. D, the migration and invasion index were quantified (n = 3). E, representative images of the wound-healing assay of PDLIM5 knockdown A549 cells with or without SMAD3 overexpression. The images were captured at 0 and 24 h after wounding. Scale bar, 200 μm. F, the wound-healing rates were analyzed by ImageJ software (n = 3). The data are shown as means ± S.D. Analysis was performed using one-way ANOVA with Tukey post hoc test for A, B, D, and F. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant.