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. 2020 Aug 31;10:14302. doi: 10.1038/s41598-020-70510-3

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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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Phenotypic myotube characterization. (A) Phase contrast images showing myotube derivation from myoblasts from the three cell lines. Scale bar = 100 μm. (B) ICC images of myotube cultures stained for MyHC and DAPI. Note the unfused background cells in the DAPI⁺/MyHC SOD1 cultures. Imaged with Volocity software, version 6.3.0 (PerkinElmer). Scale bar = 50 μm. (C) Bar graph showing slightly decreased myotube cross sectional width compared in SOD1 cultures relative to WT (Error bars CI = 90%). (D) Average myotube thickness **p value = 0.018. (E) Box and Whisker plots describing the lengths of WT and SOD1 myotubes. ***p value < 0.001. One-way ANOVA, followed by Dunnett’s test. (F) Bar graph showing fewer fusion events in the SOD1-mutant cultures compared to WT, evidenced by fusion index. (G) Bar graphs indicating lower average myotube number per area in SOD1 cultures compared to WT. Error bars: SEM *p < 0.1; **p < 0.01; ***p < 0.001 by Student T-test, 2 tails, unequal variance.