Fig. 3

High sensitivity of TGF-β1-induced fibrotic mediators in fibrotic lung fibroblasts. Subconfluent fibroblasts from 12 controls and 12 patients with PF were cultured in serum-free (SF)-DMEM for 24 h and then incubated in the presence or absence of TGF-β1 (0.25 ng/mL) and/or pirfenidone (100 ng/mL) for 48 h. Proteins from monolayer cultured fibroblasts were extracted and subjected to western blot analysis, and media were harvested from monolayer cultures and evaluated for CTHRC1 and TGF-β1 levels using immunoassay. Expression of (a) CTHRC1 (30 kDa) and (b) FHL2 (30 kDa) from fibroblasts isolated from controls and patients with PF in the presence or absence of TGF-β1 (0.25 ng/mL). Vertical axis: Protein expression normalized to β-actin expression. Immunoassay of (c) CTHRC1 and (d) TGF-β1. Vertical axis: mediator production expressed as an amount. Symbols represent the mean values for individual patients, as assessed in two separate experiments. Horizontal axis: conditions. Student’s paired t test was used. *P < 0.05, **P < 0.01, ***P < 0.001