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. 2022 Jan 7;10:3. doi: 10.1038/s41413-021-00171-z

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© The Author(s) 2022

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 — KDM6B is critical for E2-mediated osteogenic differentiation of DMSCs. a Depletion of KDM6B in DMSCs via two different shRNAs (sh1KDM6B and sh3KDM6B) by qRT-PCR. b Alkaline phosphatase staining and quantification of KDM6B-depleted DMSCs after 5 days in osteogenic media with and without E2. c Alizarin Red S staining and quantification of KDM6B-depleted DMSCs after 14 days in osteogenic media with and without E2. d qRT-PCR of osteogenic genes (HOXC6, DLX5, ALP, OCN) in the KDM6B-depleted DMSCs following osteogenic media induction with and without E2. e Overexpression of KDM6B in KDM6B-depleted DMSCs confirmed by western blot analysis. f Alkaline phosphatase staining and quantification of DMSC/Scrsh/V, DMSC/shKDM6B/V, and DMSC/shKDM6B/Flag-KDM6B. g Alizarin Red S staining and quantification of DMSC/Scrsh/V, DMSC/shKDM6B/V, and DMSC/shKDM6B/Flag-KDM6B. Data are presented as the mean ± SEM (n = 3). For a and d, data are shown as fold expression of target genes after normalization to DMSC/Scrsh. For a, the groups with shKDM6B were compared with Scrsh by using one-way ANOVA with Tukey’s post hoc test. For b, c, d, f and g, all the groups were compared by two-way ANOVA with a Bonferroni post hoc test. Asterisks were assigned to P values with statistical significance (*P < 0.05; **P < 0.01; ***P < 0.001)