Fig. 4. Effects of BMP-3 and -4 on chemotaxis and gel contraction and BMP-3 and -4 regulation in fibroblasts from control and lung fibrotic (interstitial pneumonia, IP) subjects. (A-D) Effect of various concentrations of BMP-3 and -4 in the presence or absence of transforming growth factor (TGF)-β1 on HLF1-mediated gel contraction (A, C) and chemotaxis (B, D). The vertical axes in (A) and (C) show gel size on day 3, expressed as a percentage of the control. The vertical axes in (B) and (D) show the number of migrated cells per five high-power fields (5HPF). (E, F) Effect of BIX02189 on TGF-β1-induced BMP-3 and -4 expression. Sub-confluent fibroblasts were cultured in serum-free (SF)-DMEM for 24 h, and then incubated with or without TGF-β1 or BIX02189 for 8 h. Total protein was then extracted, and western blotting was performed with antibodies against BMP-3 and -4. The vertical axes in (E) and (F) show the relative intensity of BMP-3 or -4 vs. β-actin ratio; the horizontal axes show the conditions. The values represent mean ± SEM of at least three independent experiments. For (A-D), ^†P<0.05, ^††P<0.01 vs. solvent control; *P<0.05, **P<0.01 vs. with stimulus. For (E, F), ^†P<0.01.