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. 2017 Oct 25;7:14061. doi: 10.1038/s41598-017-14444-3

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© The Author(s) 2017

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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Gene expression of C5aR1 and C5aR2 in primary osteoblasts (OB) and osteoclasts (OC) (a). Representative images of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in spine (b), arrowheads indicate osteoclasts, scale bar 100 µm. In vitro osteogenic differentiation and proliferation of primary osteoblasts and osteoclasts isolated from long bones (c–g). Quantitative analysis of TRAP-positive multi-nucleated cells (MNC) (c), n = 12-20, ***p < 0.001. Staining for TRAP in bone marrow-isolated osteoclast progenitor cells after differentiation (d), arrowheads indicate osteoclasts, scale bar 100 µm. TUNEL staining in bone marrow-isolated osteoclast progenitor cells after differentiation (e), scale bar 50 µm. BrdU proliferation assay of primary osteoblasts (f), n = 4–7. Expression of osteogenic marker genes in primary osteoblasts after 14 days of differentiation (g), n = 5–7, data were not significantly different, *p < 0.05.