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. 2023 Apr 7;8:142. doi: 10.1038/s41392-023-01354-2

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© The Author(s) 2023

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. 2 — Inactivation of OTUB1 in osteoblast cells leads to bone mass loss. a Representative micro-CT images of whole femoral (top) and trabecular (bottom) bones from 4- and 8-week-old male OTUB1 CTRL and CKO mice. n = 5 per group. Scale bars, 0.5 mm. b Histomorphometric analysis of trabecular bones from (a), including bone mass density (BMD), bone volume per tissue volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular spacing (Tb.Sp) and cortical density. n = 5 per group. c Serum levels of procollagen type 1 amino-terminal propeptide (P1NP) from 8-week-old male OTUB1 CTRL and CKO mice. n = 6 per group. d H&E staining and quantitative analysis of osteoblast numbers (N.Ob/BS) in femurs from 8-week-old male Otub1 CTRL and CKO mice. n = 4 per group. Scale bar, 100 μm. e Representative images of Von Kossa staining of femurs from 8-week-old male OTUB1 CTRL and CKO mice. n = 3 per group. Scale bar, 100 μm. f Representative images of calcein double staining and quantitative analysis of mineralization apposition rate (MAR) of the trabecular bones (TB) and cortical bones (CB) from 8-week-old male OTUB1 CTRL and CKO mice. n = 3 per group. Scale bar, 20 μm. g Immunofluorescence assay for expression of osteoblast-differentiation-related marker genes, including Osx and Col1a1 in OTUB1 CTRL and CKO mice. n = 3 per group. Scale bars, 200 μm. h Quantitative RT-PCR analysis of osteogenesis genes mRNA levels in 8-week-old male OTUB1 CTRL and CKO mice. n = 6 per group. i Representative images of ALP and ARS staining of BMSCs from OTUB1 CTRL and CKO mice after cultured in osteogenic medium for 14 and 28 days. *p < 0.05, **p < 0.01, ****p < 0.0001, n.s., not significant. All data are shown as the mean ± SEM