Figure 4.

E7130 reduced TGF-β-induced α-SMA expression in the in vitro CAF-inducing system. (a) BJ cells (normal human lung fibroblasts) were co-cultured with FaDu cells for three days in the absence (vehicle) or presence of A83-01, a potent selective TGF-β-R inhibitor, and the expression of α-SMA was analysed by immunofluorescence staining (red colour). Samples were also stained with anti-pan-human cytokeratin (green colour) for cancer cell staining and DAPI (blue colour) for nuclear staining. b-f, BJ cells were treated with E7130 (0.15 nmol/L in immunofluorescence staining data and indicated concentrations in western blot analysis data) and TGF-β (1 ng/mL) for 2 days. (b–e) Samples were stained with the indicated antibody (red colour) and DAPI (blue colour) for nuclear staining. (f) Western blot images and quantification of the images. The graph shows ratios of the results of treatment groups to the non-treated group. (g,h) BJ cells were pretreated with defactinib (1 μmol/L) and TGF-β (1 ng/mL) for 2 days. The lens magnification used was ×4 (a–c,g,h) or ×40 (d,e).