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. 2022 Jan 24;28(1):104–116. doi: 10.1038/s41591-021-01615-z

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© The Author(s) 2022

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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Extended Data Fig. 5 — (a) Numbers of ChAT positive motor neurons at lumbar levels 4 and 5 in WT/WT (black) and MN-Δ14/WT (c14/WT; ChAT-Cre and Δ14/WT, purple and blue respectively) animals normalized to the wild type controls. (b) Numbers of ChAT positive motor neurons at lumbar levels 4 and 5 in WT/WT (black) and MN-Δ14/Δ14 (c14/c14; ChAT-Cre, Δ14/c14; ChAT-Cre, and Δ14/Δ14, red, green, and brown respectively) animals normalized to the controls. (c) Density of Iba1-positive microglial cells at lumbar levels 4 and 5 in WT/WT (black) and MN-Δ14/WT (c14/WT; ChAT-Cre and Δ14/WT, purple and blue respectively) animals. (d) Density of Iba1-positive microglial cells at lumbar levels 4 and 5 in WT/WT (black) and MN-Δ14/Δ14 (c14/c14; ChAT-Cre, Δ14/c14; ChAT-Cre, and Δ14/Δ14, red, green, and brown respectively) animals. (e) Percentages of NMJs classified as fully innervated (green), partially denervated (yellow), and fully denervated (red) in the tibialis anterior (left) and soleus (right) muscles in CTRL (FUS WT-expressing control) and MN-P517L/Δ14 (FUS P517L/c14; ChAT-Cre) animals. *P < 0.05, **P < 0.01 and ***P < 0.001 indicates significant differences in fully denervated NMJs using two-way ANOVA with Sidak´s post hoc test. Data shown as mean ± SD. N = 3 animals per group. (f) Density of Iba1-positive microglial cells at lumbar levels 4 and 5 in CTRL (FUS WT-expressing control, black) and MN-P517L/Δ14 (FUS P517L/c14; ChAT-Cre, green) animals. (g) Percentage of cross-sectional area that is GFAP-positive at lumbar levels 4 and 5 in CTRL (FUS WT-expressing control, black) and MN-P517L/Δ14 (FUS P517L/c14; ChAT-Cre, green) animals. (h) Normalized grip strength of forelimbs (left) and hindlimbs (right) in CTRL (FUS WT-expressing control, black) and MN-P517L/Δ14 (P517L/c14; ChAT-Cre, green) animals normalized to the controls. *P < 0.05, **P < 0.01 and ***P < 0.001 using two-way ANOVA with Sidak´s post hoc test. Data shown as mean ± SD. N = 5–28 per group at different time points. For a-d, *P < 0.05, **P < 0.01 and ***P < 0.001 using two-way ANOVA with Tukey´s post hoc test. Data shown as mean ± SD. N = 3 animals per genotype. For f and g, *P < 0.05, **P < 0.01 and ***P < 0.001 using two-way ANOVA with Sidak´s post hoc test. Data shown as mean ± SD. N = 3 animals per group. CTRL=littermate c14/WT and/or c14/c14 animals depending on the breeding scheme.