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. 2020 Mar 11;10:4524. doi: 10.1038/s41598-020-61347-x

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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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Effect of various laminins on AChR cluster formation and remodeling in human primary myotubes. (a) Quantification of AChR clusters formed by human primary myotubes that were cultured on the indicated laminins. (b) Quantification of plaque-shaped AChR clusters formed by human primary myotubes that were cultured on the indicated laminins, presented as the average percentage of clusters. (c) Example of plaque-shaped cluster. (d) Quantification of perforated clusters that contained synaptic podosomes, presented as the average percentage of clusters. (e, f) Examples of clusters that contained actin-rich podosomes that were immunostained for podosome cortex protein markers vinculin (e) and LL5β (f). Scale bar = 6 μm. Quantifications represent the average number of clusters. For all laminins except laminin-221, the total number of clusters that were used for the analysis was> 140. For laminin-221, 83 clusters were used. Clusters were collected from three independent experiments performed on three different days. *p < 0.05, **p < 0.005, ***p < 0.0005. For statistical analysis we used one way ANOVA with Dunnett test; laminin-111 used as a reference. Error bars represent SEM values. AChR (red) was visualized with α-bungarotoxin. F-actin (blue) was visualized with phalloidin.