Figure 5.

Effect of matrix metalloproteinase (MMP) 19 catalytic domain on A549 in vitro phenotypes: epithelial–mesenchymal transition, migration, and invasion. (A) Schematic representation of the E213A MMP19 point mutation in the catalytic domain. (B) Pitt cohort of MMP19 expression in protein in A549 cells transfected with wild-type (WT) and the E213A mutated MMP19 construct by immunoblotting. (C) Quantitative reverse-transcriptase polymerase chain reaction was performed for markers of epithelial–mesenchymal transition. Data represent the mean ± SEM (*P < 0.05 compared with mock transfection control). (D) Immunofluorescent staining of mesenchymal markers (anti-ACTA2 and anti-VIM, green) and epithelial marker (anti-CDH1, red). Cells were counterstained with DAPI (blue). (E) Induction of A549 cell migration and invasion with expression of MMP19 WT and MMP19E213A in matrigel. Data represent mean of five experiments with SEM. Five high-power fields were counted for each experiment (*P < 0.05 compared with mock transfection control). (F) Scratch assay was performed on A549 following transfection of the mock control, MMP19 WT, and the E213A MMP mutant. (G) Quantification of the gap coverage as described in the Methods section (*P < 0.05, compared with time = 0 control). ACTA2 = α-smooth muscle actin; CDH1 = E-cadherin; VIM = vimentin.