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. 2019 Aug 20;9:12094. doi: 10.1038/s41598-019-48580-9

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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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SM decreased Akt activity in human eSCs. (A–D) Human eSCs were incubated either under terrestrial gravity (1 g) or under SM for 36 h, lysed, and analyzed by western blotting (A,C). Western blot images were analyzed using ImageJ to determine the phosphorylation of pS473-Akt relative to Akt, pT308-Akt relative to Akt, pT389-S6K1 relative to S6K1, pS65-4EBP1 relative to 4EBP1, and pT47/36-4EBP1 relative to 4EBP1 (B), and the expression of mTOR, raptor, and rictor relative to tubulin (D). The cells were treated as in (A), subjected to immunoprecipitation using antibodies against mTOR, and analyzed by western blotting. Abbreviations: simulated microgravity (SM). Data are expressed as mean ± SD, with paired t-tests performed as indicated. *P < 0.05, **P < 0.01 versus control at each indicated time.