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. 2012 Dec 5;304(4):C289–C298. doi: 10.1152/ajpcell.00298.2012

Fig. 1.

Fig. 1.

Transforming growth factor-β (TGF-β) induced smooth muscle cell (SMC)-specific gene expression in human embryonic stem cell-derived mesenchymal cells (hES-MCs). A: TGF-β stimulates SMC marker mRNA expression in a dose-dependent manner. hES-MCs were treated with various dosages of TGF-β as indicated for 24 h. mRNA expression of SMC markers was examined by qPCR. *P < 0.05 compared with the untreated group for each individual marker. B: TGF-β stimulates SMC marker mRNA expression in a time-dependent manner. hES-MCs were treated with 1 ng/ml TGF-β for various time points as indicated. qPCR was performed as in A. *P < 0.05 compared with the untreated group for each individual marker. C: TGF-β induces SMC marker protein expression. hES-MCs were treated with 1 ng/ml TGF-β for various time points as indicated, and Western blot analysis was performed. α-TUBA was an internal control. D: global expression of SMC-specific genes in TGF-β-treated hES-MCs. hES-MCs were treated with 1 ng/ml TGF-β for 0 (D0), 1 (D1), 2 (D2), and 3 days (D3) to detect SMC marker expression by immunostaining. E: percentage of smooth muscle α-actin (α-SMA)-positive cells before and after 3 days of TGF-β treatment. *P < 0.05 compared with untreated cells (D0). F: stability of SMC phenotype after trypsinization and reculture. hES-MCs were treated with vehicle (-β) or 1 ng/ml TGF-β (+β) for 3 days. Cells were then trypsinized and recultured without TGF-β (+β-β). After 2 days of incubation, cells were trypsinized again and recultured without TGF-β for an additional 2 days (+β-β-β). Marker expression was examined by Western blotting.