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. 2021 Jun 3;11:11695. doi: 10.1038/s41598-021-91203-5

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Co-localisation of pre- and post-synaptic membrane proteins, motor nerve axons and myosin heavy chain positive fibres indicate neuromuscular junction formation in engineered tissues. (a) Confocal tile scan of neuromuscular tissue detailing AChR (left) and SV-2 (middle), with overlay (right) evidencing multiple synaptic contacts (outlined—white boxes) via co-localisation of pre- and post-synaptic markers and Tuj-1 + neuronal axons. (b) Confocal tile scan of neuromuscular tissues evidencing AChR, Tuj-1 + nerve terminal and myosin heavy chain co-localisation. (c) Zoom of synaptic contacts highlighted in (b) via white box outline. Images were collected across n = 8 neuromuscular tissues. Scale bars = (a, b) 100 µm, (c) 50 µm.