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. 2020 May 19;10:8239. doi: 10.1038/s41598-020-65067-0

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© The Author(s) 2020

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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Differentiation characteristics of developmental, mature, and aged SMSCs. (A) Representative images of developmental, mature, and aged SMSCs (MyoD⁺ cells) under growth-stimulating conditions for 48 h. Arrows in (A) indicate fusion of ≥3 MyoD⁺ nuclei (nascent myotube formation). (B–D) Quantitative analyses of (B) number of nascent myotubes, (C) fusion index, and (D) MHC protein expression from developmental, mature, and aged SMSCs (n = 3 SMSCs isolated from different mice/growth stage). (E) Representative images of MHC staining in developmental, mature, and aged SMSCs under growth-stimulating conditions for 48 h. Arrows in (E) indicate nascent myotubes (MHC⁺) that contains ≥3 nuclei fusion. *p < 0.05 and **p < 0.01 compared to developmental stage and ^#p < 0.01 compared to ageing stage. Images were taken at ×200 magnification.