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. 2020 Jan 30;4(3):e10338. doi: 10.1002/jbm4.10338

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© 2020 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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In vitro osteoclast‐precursor (OCP) cell differentiation assays in germ‐free versus segmented filamentous bacteria‐monoassociated mice. (A–D) Day 4 OCP culture tartrate‐resistant acid phosphatase stain assay (n = 4 to 5/gp). (A) Representative images (×100) of CD11b^neg OCP cultures stimulated with treatment (CSF1 & RANKL) media for 4 days. (B) N.Oc = number of osteoclasts enumerated within four fields of view per well. (C) Oc.Ar/Oc = average osteoclast area. (D) N.Nc/Oc = nuclei number per osteoclast. (E–H) Day 6 OCP culture TRAP stain assay (n = 4 to 5/gp). (E) Representative images (×100) of CD11b^neg OCP cultures stimulated with treatment (CSF1 & RANKL) media for 6 days. (F) N.Oc. (G) Oc.Ar/Oc. (H) N.Nc/Oc. Unpaired t test; data are presented as mean ± SEM, *p < 0.050, **p < 0.010.