Figure 2 | Scientific Reports

Figure 2

From: Direct phenotypic conversion of human fibroblasts into functional osteoblasts triggered by a blockade of the transforming growth factor-β signal

Figure 2

An addition of ALK5 i II inhibited smad2/3 signaling in fibroblasts. (A) HDFs were seeded onto 60-mm dishes, and on the next day culture supernatant was replaced by fresh complete medium with/without the indicated concentrations of TGF-β inhibitors. After culturing for 3 days, culture supernatant was replaced by fresh one, and cells were lysed 30 min later. ELISA was performed to evaluate relative ratio of phospho-smad2/3 per total smad2/3 (the value for untreated HDFs was set to 1). Values are means ± S.D. (n = 3). *P < 0.05, **P < 0.01 vs. untreated control. #P < 0.05, ##P < 0.05 vs. HDFs treated with ALK5i II at the same concentration. (B and C) HDFs were seeded onto 60-mm dishes, and on the next day culture supernatant was replaced by fresh complete or osteogenic medium with/without ALK5 i II as indicated. After culturing for 3 days, culture supernatant was replaced by fresh one, and cells were lysed 30 min later. Western blotting analyses were performed using the indicated antibodies.

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