Fig. 5

Effects of pirfenidone on CTHRC1-mediated regulation in lung fibroblasts. Subconfluent HFL-1 cells were cultured in SF-DMEM for 24 h and then incubated in the presence or absence of different concentrations of rhCTHRC1. Western blot analysis of the effects of different concentrations of rhCTHRC1 on targets related to fibrotic processes (a). Effects of different concentrations of rhCTHRC1 on HFL-1 cell-mediated collagen gel contraction (b) and chemotaxis (c). Effects of different concentrations of rhCTHRC1-mediated targets assayed using western blot analysis (d-h). The vertical axis shows the relative intensities of FHL2 (D), α-SMA (e), fibronectin (f), BMP4 (g), Gremlin1 (h) versus β-actin; the horizontal axis shows the conditions. Subconfluent HFL-1 cells were cultured in SF-DMEM for 24 h and then incubated in the presence or absence of rhCTHRC1 (100 ng/mL) and pirfenidone (100 ng/mL) for 48 h. Western blot analysis to analyze the effects of pirfenidone on rhCTHRC1-mediated targets related to fibrotic processes (i). Effects of pirfenidone on rhCTHRC1-mediated collagen gel contraction (j) and chemotaxis (k). Effects of pirfenidone on the expression levels of rhCTHRC1-mediated targets assayed using western blot analysis (l-p). The vertical axis shows the relative intensities of α-SMA (l), fibronectin (m), FHL2 (n), BMP4 (o), Gremlin1 (p) versus β-actin; the horizontal axis shows the conditions. Collagen gel contraction, vertical axis: gel size measured after 2 days of contraction expressed as a percentage of the initial value. Chemotaxis, vertical axis: number of migrated cells per 5 HPF. Horizontal axis: conditions. Values represent means ± SEMs of three to five separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001