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. 2019 May 20;10:2226. doi: 10.1038/s41467-019-10197-x

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© The Author(s) 2019

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 — PTEN overexpression decreases adipogenesis and increases osteogenesis in BM-MSCs. a–e BM-MSCs were transfected with a control plasmid pcDNA6^TMV5-HisA (CTR) or an overexpression plasmid, pcDNA6-PTEN (PTEN O/E), followed by induction for b, c adipogenic or d, e osteogenic differentiation. a Cells were analyzed by western blotting. b Cells induced for adipogenic induction were stained by nile red and (right) quantification of nile red positive cell percentages. Bar = 25 μm. c Quantitative RT-PCR for the mRNA levels 7 days after differentiation. d Cells induced for osteogenic differentiation were analyzed by Arsenazo III kit and (right) quantification of the calcium contents relative to DNA contents at 2 weeks. Bar = 200 μm. e Quantitative RT-PCR for the mRNA levels 7 days after differentiation. c, e Results are shown as the relative expression to GAPDH (mean ± SD of three independent experiments), and significance was determined by Student’s t-test. (*p < 0.05 and **p < 0.01 versus CTR)