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. 2022 Oct 6;13:5883. doi: 10.1038/s41467-022-32915-8

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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. 9 — a Schematic representation of the active DNA demethylation cycle and experimental design for the shown experiments in differentiating osteoblasts. b Dot blots at day 14 with and without VitC and/or the DNMT-inhibitor RG108, which decreases global CpG methylation levels (5mC). c ECM deposition, alkaline phosphatase activity (ALPL) and ECM mineralization in the presence or absence of VitC and/or RG108. d Dot blots for 5hmC and 5fC (formyl-cytosine) in osteoblasts at indicated time points in the presence or absence of VitC. e Osteoblastic gene expression after VitC and/or RG108 administration. Bar graphs represent mean ± SD; *p < 0.05; **p < 0.01; ***p < 0.001. Two-way ANOVA analysis with Sidak’s multiple comparison tests (c and e), groups compared to day 0 Ctrl or as marked in e. In c, for ECM RG108- VitC- vs RG108 + VitC- **p = 0.0062 and RG108- VitC+ vs RG108 + VitC + **p = 0.0029. N = 3 (c, e) per group, from biologically independent experiments. Source data as well as exact p = values for all comparisons in (e) are provided in the Source Data File.