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. 2021 Sep 13;4:1066. doi: 10.1038/s42003-021-02504-4

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© The Author(s) 2021

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. 3 — a Diagram showing an experimental setup for ex vivo deletion of Fbxw7. Isolation of MSCs from the bone marrow of UbcCre^ERT2; Fbxw7^f/f mice (1) was followed by 4OHT-induced deletion of Fbxw7 (2), osteogenic induction (3) and analysis of osteoblast differentiation (4). b–e Osteoblast differentiation of UbcCre^ERT2; Fbxw7^f/f-derived MSCs treated with mock or 4OHT before osteogenic induction (n = 3 different mice). Differentiation was measured at 14, 21, and 28 days after osteogenic induction via Alizarin Red S staining (b) and mRNA levels of osteoblast differentiation markers Runx2, Osterix (Osx), Osteocalcin (Ocn), Osteopontin (Opnt) and Alkaline phosphatase (Alp) at 14 (c), 21 (d), and 28 days (e) after osteogenic induction. mRNA levels of Fbxw7 were analyzed to confirm deletion of the gene. Images were taken from 96-well plates. Well diameter: 5 mm (b). Data are presented as means ± SEM. Student’s t-test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.