Fig. 3

PDBu and HGF promoted cortactin expression, cell migration and invadopodia formation in NSCLC. a A549 and H1299 cells were treated with or without HGF (20 ng/mL) or PDBu (500 nM), and invadopodia were visualized by colocalization of cortactin staining with FITC (green), F-actin with phalloidin (red) and nuclei with DAPI (blue). representative images are displayed. Scale bar, 50 μm, 40× magnification. b Quantification of cells with invadopodia. c A549 and H1299 cells treated with different concentrations of HGF for 6 h or PDBu for 30 min; Cells with invadopodia were quantified. d Cell lysates from A549 and H1299 cells treated with 20 ng/mL HGF or 500 nM PDBu after blotting with cortactin antibody. e Wound healing assays in A549 cells were performed after treatment with 20 ng/mL HGF for 6 h or 500 nM PDBu for 30 min. Representative images are displayed. Scale bar,800 μm, 4× magnification, *P < 0.05. f Transwell migration and invasion assays of A549 cells were performed after treatment with 20 ng/mL HGF for 6 h or 500 nM PDBu for 30 min. Representative figures of the migrated and invaded stained cells are shown. Scale bar, 200 μm, 20× magnification. Data are shown as the mean ± SD, *P < 0.05, **P < 0.01. g Following treatment with 0, 10, and 20 μM PP2 for 24 h, A549 and H1299 cells were harvested. Levels of CTTN and the phosphorylation of CTTN were determined using Western blot analysis. h A549 and H1299 cells treated with 10 μm PP2 or 20 μm PP2 for 24 h, invadopodia were visualized as above. Representative images are displayed. Scale bar, 50 μm, 40× magnification, **P < 0.01