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. 2022 Sep 26;13:5628. doi: 10.1038/s41467-022-33184-1

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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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Fig. 1 — a Schematics depicting the experimental design for snRNA-seq, showing the injured thoracic cord and lumbar cord (with dark red representing the lesion) as well as nuclei isolation from the intact lumbar cord followed by droplet-based barcoding for single nucleus RNA sequencing. b Top, an overview of experimental design for injury and tissue collection. The lumbar spinal cord of three animals from each time point: uninjured, 1 dpi (day post injury), 1 wpi (week post injury), 3 wpi, and 6 wpi. c Uniform manifold approximation and projection (UMAP) visualization of 67,903 nuclei from uninjured and injured lumbar spinal cords, revealing 9 classes and 39 subtypes. Colored by green (neurons), yellow, astrocytes, orange-red (microglia), purple (OPCs), blue (oligodendrocytes), light blue (Schwann), light pink (pericytes), pink (ependymal), magenta (leptomeninges), brick-red (endothelial). d Multiplex immunohistochemistry (IHC) of the lumbar spinal cord from uninjured, 1 wpi and 3 wpi. Tissue was stained for NeuN (green), GFAP (yellow), IBA1 (red), TMEM119 (dark orange), and OLIG2 (blue). Scale bars are 200 µm (main) and 50 µm (inset), respectively. e Quantification of the proportion of cell types from the snRNA-seq data and immunohistochemistry in tissue. Mean ± SEM; snRNAseq, N = 3; immunohistochemistry, N = 4. Source data are provided as a Source Data file.