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. 2017 Aug 1;7:7002. doi: 10.1038/s41598-017-07633-7

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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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Synergistic effect of OA and BMP2 on MSC osteogenic differentiation. (A) ALP staining showed a significant increase of ALP activity in MSCs treated with either OA or BMP2, and this increase is further enhanced by the combined treatment at day 7. (B) Real time RT-PCR analysis reveals that expression of both early and later osteogenic markers (ALP, Runx2, OC, OPN) were significantly enhanced by combined treatment with OA and BMP2 at day 7 when compared with either OA or BMP2 alone treated MSCs. PCR data is means ± s.d. of three independent experiments performed in duplicate and all the results were normalized to control (*p < 0.05 compared with control MSCs (Co) without treatment). (C) RT-PCR data shows treatment of OA in MSCs did not cause a significant change in gene expression of phosphor-smad1/5/8 at day 7. Data is the means ± s.d. of three independent experiments. (*p < 0.05 compared with control MSCs). (D) Western Blot shows phosphor-smad1/5/8 (p-smad1/5/8) protein levels were not affected by OA treatment in MSC culture at day 7. β-actin was used as a loading control. Full-length blots/gels are presented in Supplementary Figure S1.