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. 2017 May 4;7:1506. doi: 10.1038/s41598-017-01698-0

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© The Author(s) 2017

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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The differentiation experiment of the osteoblast-like cells was done with cultivation medium (CM), differentiation medium (DM) and DM with low (17 µg/ml) and high (133 µg/ml) AMP2. After 21 days alkaline phosphatase (AP) activity assay and staining (A) and alizarin red staining and quantification (B) were done (n = 6, cells from two donors, triplicates). Results were normalized to 10⁴ cells and expressed in percentage to control (DM) which was set 100%. Data are shown as median with 25 and 75 percentiles. Kruskal-Wallis, Mann-Whitney, Bonferroni-Holm post-hoc test: *Significant difference compared to the DM group, p ≤ 0.05. (C) Gene expression at days 0, 7, 14 and 21. Results are calculated with the ΔΔCt method with efficiency correction and normalization to the control (day 0 of differentiation) and 18 S ribosomal RNA used as a reference gene (n = 2). Data are shown as means with standard deviation.