A Novel OsteomiRs Expression Signature for Osteoblast Differentiation of Human Amniotic Membrane-Derived Mesenchymal Stem Cells

Figure 6

MiR-204 inhibits the osteogenic differentiation of AM-hMSCs by targeting TGF-βR2. (a) Relative expression of miR-204 after transfection of RNA mimics in AM-hMSCs at 13 days after dexamethasone-induced osteoblastic differentiation. (b) Quantification of Alizarin Red staining for calcium deposition (bottom images) in nontransfected AM-hMSCs (control) or transfected cells with miR-204 mimics. (c) Schematic representation of p-miR report construct containing the 3′UTR of TGF-βR2 gene cloned downstream of the firefly luciferase gene. Seed sequence is indicated in blue shaded box. Point mutations in the miR-204 binding site of 3′UTR of TGF-βR2 gene are denoted in bold. (d) Luciferase assays in AM-hMSCs cotransfected with miR-204 mimics and the constructs described in (c). Nontransfected and scramble transfected cells were used as controls. Data represent the mean ± SD of three independent experiments (p<0.001; p<0.05).

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