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. 2018 Aug 14;6:24. doi: 10.1038/s41413-018-0026-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — R-spondin-2 null mesenchymal progenitor cells from Footless mutant mice show reduced osteoblastogenesis. a Alizarin red S staining of representative wells from in vitro osteogenesis assay after 10 days. b Overall quantification of Alizarin Red S staining of osteogenesis at 10 days, as shown in a. c Quantification of osteogenesis at 15 days with addition of recombinant hRSPO2 (10 nmol·L^-1). d Alkaline Phosphatase (ALP) staining of similar osteogenesis assay at 5 days. e Relative Runx2 expression after 5 days of osteogenesis in vitro. f Relative Osterix expression after 5 days of osteogenesis in vitro. g Relative Osteocalcin expression after 10 days of osteogenesis in vitro. h Relative bone sialoprotein (BSP) expression after 5 days of osteogenesis in vitro. (n = 3 cell lines from individual animals, assay performed in triplicate for each cell line (sex of pre-natal pups not determined), * indicates P< 0.05. MM maintenance media, OPM osteopermissive media with β-glycerophosphate and ascorbic acid-2-phosphate, OGM osteogenic media with OPM and 2.5 nmol·L^-1 rhBMP-6