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. 2017 Aug 8;7:7518. doi: 10.1038/s41598-017-07932-z

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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 role of Trib3 induced by phenamil in the BMP2-mediated osteogenesis and pro-adipogenesis by examining Trib3 knockdown hBMSCs. (a–d) Blocking Trib3 expression led to the loss of synergistic effect of phenamil (20 µM) on BMP2 (100 ng/mL)-based osteogenesis by the detection of ALP staining/activity (scale bar = 200 µm) (a,b), and Alizarin red S staining/quantification (scale bar = 100 µm) (c,d); (e,f) Down-regulated Trib3 attenuated the anti-adipogenic capacity of phenamil (10 µM) on high dose BMP2 (300 ng/mL)-induced adipogenesis via Adipored assay at day 7 (e) and Oil red staining (scale bar = 100 µm) at day 14 (f); (g) Phenamil exerted anti-adipogenesis via up-regulating Trb3 that inhibited PPARγ expression. (h) Trib3 knockdown attenuated phenamil (10 μM)-mediated inhibitory BMP2 (300 ng/mL)-induced adipogenic genes: PPARγ in hBMSCs. Phe, phenamil; BMP, BMP2; OM, osteogenic medium; AD, adipogenic medium. Data presented as means ± SD (n = 3/group); *p < 0.05. Uncropped western blot gels are displayed in Supplementary Figure 6.