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. 2009 Dec 3;285(7):4975–4982. doi: 10.1074/jbc.M109.035105

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Loss of Tgfbr2 in CNCC results in altered osteogenic differentiation during intramembranous ossification. A, quantitative RT-PCR analyses of indicated genes in Tgfbr2^fl/fl (open columns) and Tgfbr2^fl/fl;Wnt1-Cre (closed columns) mice at E13.5. Ocn, osteocalcin; On, osteonectin; Spp1, osteopontin; Osx, Osterix; Alp, alkaline phosphatase; ColI, type I collagen. *, p < 0.05. B, immunological analysis of osteocalcin (Ocn), osteonectin (On), and osteopontin (Opn) in Tgfbr2^fl/fl and Wnt1-Cre;Tgfbr2^fl/fl maxilla at E12.5, E13.5, and E14.5. Data shown are representative of three separate experiments. C, plot shows the ratios between osteocalcin (Ocn), osteonectin (On), and osteopontin (Opn) versus GAPDH based on quantitative densitometry of immunoblotting data in B; *, p < 0.05. Tgfbr2^fl/fl, white bars; Tgfbr2^fl/fl;Wnt1-Cre, black bars. D, osteogenic differentiation of Tgfbr2^fl/fl and Tgfbr2^fl/fl;Wnt1-Cre primary MEMM cells cultured for 14 days in osteogenic induction medium. Alkaline phosphatase (ALP) staining of Tgfbr2^fl/fl and Tgfbr2^fl/fl;Wnt1-Cre MEMM cells cultured without (Dif. −) or with (Dif. +) osteogenic inducer for 2 weeks. E, mRNA expression of indicated genes after no osteogenic induction (−) or induction (+) in Tgfbr2^fl/fl and Tgfbr2^fl/fl;Wnt1-Cre MEMM cells. Data shown are representative of three separate experiments. F, graph shows quantitative densitometry analysis of gel electrophoresis data in E. Data shown are representative of three separate experiments. Lane 1, Tgfbr2^fl/fl MEMM cells without osteogenic induction; lane 2, Wnt1-Cre;Tgfbr2^fl/fl MEMM cells without osteogenic induction; lane 3, Tgfbr2^fl/fl MEMM cells with osteogenic induction; lane 4, Wnt1-Cre;Tgfbr2^fl/fl MEMM cells with osteogenic induction.