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. 2017 Mar 7;18(3):564. doi: 10.3390/ijms18030564

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© 2017 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Rspo1 neutralizing antibody (Rspo1 Ab) (1 μg/mL) abolished the enhanced osteogenic differentiation and Wnt/β-catenin activity by CMS (5% elongation) in BMSCs. (A) qRT-PCR analysis of osteogenic differentiation markers Ocn, Col-1a1 and ALP (Day 3); (B) relative ALP activities (Day 7); (C) ELISA measurement for supernatant Ocn (Day 3); (D) Representative images of ALP staining (Day 7) and Alizarin red staining (Day 14); (E) qRT-PCR analysis of the Wnt target genes Axin2 and Tcf1; (F) Western blot analysis of the intranuclear level of β-catenin and (G) Topflash dual-luciferase activity assay of BMSCs after 24 h of osteogenic differentiation. The luciferase activities were presented as fold changes normalized to Renilla compared to Con group cells. All data were confirmed by three repeated tests and presented as the mean ± SD, * p < 0.05. p-Values are based on ANOVA and post hoc analysis.