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Fig. 3. Lack of Kcnma1 decreases the capacity of osteoblast differentiation. (A and B) Expression and quantitative analysis of osteogenic markers, including Runx2 and Col1a1, by western blot. Quantification of protein levels was normalized to GAPDH. (C) qRT-PCR analysis of Alp and Runx2 in long bones from Osterix-Cre(–); Kcnma1f/f and Osterix-Cre(+); Kcnma1f/f mice. (D) The ratio of Rankl/Opg were determined in the bone tissues of Osterix-Cre(–); Kcnma1f/f and Osterix-Cre(+); Kcnma1f/f mice by RT-PCR. (E) Representative images of TRAP staining in the trabecular region of the tibiae. Scale bars = 100 μm. (F) ALP staining of primary osteoblasts isolated from the indicated mice after 14 days of culture. Scale bars = 100 μm. (G) Representative images of Alizarin Red staining of osteoblasts cultured in DMEM supplemented with 10 mM β-glycerophosphate and 50 μg/ml ascorbic acid. Scale bars = 100 μm. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Mol. Cells 2021;44:557~568
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