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. 2020 Nov 5;11:5596. doi: 10.1038/s41467-020-19360-1

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© The Author(s) 2020

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. 4 — a Intracellular αKG concentration in MSCs isolated from young (3-month-old) and aged (18-month-old) mice (n = 3, by two-tailed Student’s t-test). b Intracellular αKG concentration in aged MSCs in response to αKG treatment for 3 days (n = 3, by one-way ANOVA with Tukey’s post hoc test). c EdU incorporation in aged MSCs treated with different concentrations of αKG (n = 4, by one-way ANOVA with Tukey’s post hoc test). αKG treatment increased the number of EdU⁺ cells. d Crystal violet staining of colony-forming units of MSCs. e Quantitative analysis of colony-forming unit assay (colonies per well, n = 6, by two-tailed Student’s t-test). f, g Scratch assay of aged MSCs and quantitative analyses. Magenta dotted lines indicate start (0 h), half (9 h), and end (18 h) positions of MSCs after scraping (n = 5, by two-way ANOVA with Sidak’s multiple comparisons test). Scale bar, 500 μm. h, i Crystal violet staining of migrated MSCs from the upper chamber to bottom lower of the membrane and quantitative results (n = 3, by two-tailed Student’s t-test). Scale bar, 100 μm. j Representative images of ALP, ARS and Van Gieson staining of aged MSCs. k Quantitative analysis of the ALP activity in MSCs (n = 3, by two-tailed Student’s t-test). l Quantitative analysis of calcium mineralization in MSCs (n = 3, by two-tailed Student’s t-test). m Quantitative RT-PCR results of mRNA expression of Runx2, Sp7, Col1a1, Alp, and Bglap in MSCs treated with vehicle/αKG (n = 3, by two-tailed Student’s t-test). Data are expressed as mean ± SEM.