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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: TGF-β1-induced differentiation of SHED into functional smooth muscle cells

Fig. 2

Characterization of SHED-derived SMCs. a Expression levels of SMC-specific marker gene expression relative to GAPDH at different induction time points. The expression levels of all SMC-specific marker genes peaked on day 5 (p < 0.05, compared with other time points). At passage 2, the gene expression levels of α-SMA and Calponin 1 declined but were still significantly higher than the control group (p < 0.05); while the gene expression level of SM22α declined to the same level as the control group (p > 0.05). * ^ Φ: p < 0.05 compared to all other time points. b The protein expression levels of SM22α and Calponin 1 were analyzed by western blot utilizing β-actin as the internal marker. Numbers depict the band density normalized against untreated controls and β-actin. c SB-431542 suppressed TGF-β1-mediated SMC differentiation. * ^ Φ: p < 0.05 between TGF-β1 group and SB-431542 + TGF-β1-treated group. d Smad2/3 became phosphorylated within 30 minutes of exposure to TGF-β1, and SB-431542 inhibited this phosphorylation. Numbers depict the band density normalized against untreated controls and β-actin. e SHED-derived SMCs were also analyzed for expression of SMC-specific markers (α-SMA, Calponin, SM22α and SM-MHC) with flow cytometry (α-SMA+ 86.1%, SM22α + 93.9%, Calponin + 56.8%, and SM-MHC+ 88.2% respectively) (the isotype control in red) and (f) immunofluorescence staining (nuclei in blue). All experiments were performed three times (N = 3). SM-MHC smooth muscle-myosin heavy chain, SHED stem cells from human exfoliated deciduous teeth, α-SMA alpha-smooth muscle actin

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