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. 2022 Jun 20;13(4):2102–2117. doi: 10.1002/jcsm.13023

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© 2022 The Authors. Journal of Cachexia, Sarcopenia and Muscle published by John Wiley & Sons Ltd on behalf of Society on Sarcopenia, Cachexia and Wasting Disorders.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Classification of nucleus/cell types in normal and denervated gastrocnemius (gas) muscles. (A) Immunofluorescent staining: Type I fibres are labelled with anti‐Myh7 antibody (red) and co‐stained with laminin (green). Images were taken at ×20 magnification. (B) Comparison of cross‐sectional area (CSA) between normal and denervated conditions. The data are presented as mean ± SEM (n = 6). (C) Fibre size distribution of normal and denervated gas muscles. The size of 300–400 fibres in muscles of each animal was assessed (n = 6). (D) Uniform manifold approximation and projection (UMAP) visualized nuclear clusters, which are coloured and labelled according to cell identities, in normal (left panel) and denervated (right panel) gas muscles. Type I, type I myonuclei; type IIa, type IIa myonuclei; type IIx, type IIx myonuclei; type IIb, type IIb myonuclei; MTJ, myotendinous junction nuclei; NMJ, neuromuscular junction nuclei; MuSCs, muscle satellite cells nuclei; FAPs, fibro‐adipogenic progenitors nuclei; macrophage, macrophages nuclei; ECs, endothelial cells nuclei; Pericytes, pericytes nuclei; adipocyte, adipocyte nuclei. (E) UMAP plot displaying the cell identity of each nuclear cluster. (F) Proportion of nuclear types in normal and denervated muscles. Each nucleus/cell type is colour‐coded. (G) UMAP plots with pseudotime trajectories of all nuclei obtained from normal and denervated muscles. The black lines on the UMAP plots represent branched trajectories. Each point denotes a single nucleus. Left panel: Nuclei are colour‐coded according to their cluster assignments in (D). Right panel: Nuclei are coloured by conditions (blue, normal; pink, denervation).