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. 2017 Apr 26;7:1177. doi: 10.1038/s41598-017-01311-4

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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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Myogenic differentiation of iMuSCs and NMJ formation. (A) Whisker plots summarize the expression of NMJ-related genes as well as the myogenic genes within the neurosepheres in culture, analyzed by qPCR. (B) After the neurospheres were replaced into monolayer culture in MD Medium, their recovered their myogenic ability, formed myotube-like structures, and begun expression of myogenic protein MyoD (green, B), MyHC (Red, B; green, C,D). The attached neurospheres also formed neuron-like structures expressing β-Tubulin III (red, D). The presence of NMJs was shown by Btx staining (red, C; yellow, D). Nuclei were stained with Dapi. Scale bar = 100 µm.