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. 2017 Jul 26;7:6640. doi: 10.1038/s41598-017-06794-9

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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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Effects of Stretch on Smooth Muscle Protein Expression. Western blots reveal that cells stretched to 10% for two 16 hour regimens expressed more SMA protein than controls (A), (p < 0.05). Cells stretched to 10% for two 16 hour regimens also expressed more SM22 protein (B), (p < 0.05). There was no statistical difference in CNN protein expression between cells stretched to 10% and their respective controls (C). Cells stretched at the maximum strain (10%) and duration (2× for 16 hours) as well as their respective control were stained with anti-calponin ((D): controls; (E): stretched MSCs) and the length of cells with a bright signal (above a specific pixel intensity threshold) was compared to that of cells with a weaker signal (F). Visualizing the cell contours up close, the majority of cells with a strong Calponin signal appeared shorter than longer appearing MSCs with a weaker Calponin signal. There was no statistical difference between the length of control cells expressing strong CNN signal or cells expressing weak amounts of protein. Stretched cells that exhibited weak expression of CNN were significantly longer than their strongly expressing counterparts (p < 0.05).