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. 2022 Apr 12;8:34. doi: 10.1038/s41421-022-00377-3

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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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Fig. 2 — a Representative flow-cytometry scatter plots to assess microglia proliferation in lumbar spinal cord after sciatic nerve injury. The proportion of proliferating Ki67 (+) microglia in Cx3cr1-creER-yfp/+; +/+ animals increased 2 and 3 days after nerve injury. b Quantification of flow cytometry highlights the time course of nerve injury-induced microglia proliferation. In control Cx3cr1-creER-yfp/+; +/+ animals, microglia proliferation started at day 2, persisted through day 3, and then gradually reduced to termination by day 7 after nerve injury. In contrast, the early phase of microglia proliferation on day 2 after nerve injury was completely abolished in Cx3cr1-creER-yfp/+; Myc fl/fl cKO animals, but the late phase and the termination phase of microglia proliferation remained intact in cKO animals. c, d Representative Ki67 IHC staining (c) and the associated quantification (d) of lumbar dorsal cord microglia after nerve injury. Animals with Myc deletion from adult microglia had significantly less Ki67 (+) proliferating microglia in lumbar dorsal cord (DC) 2 days, but not 3 days, after nerve injury. The arrowheads point to the Ki67 (+) microglia, and the dashed lines outline the dorsal lumbar cord. Microglia are identified by YFP staining. e Differential expression volcano plot of pseudobulk scRNA-Seq analysis. The G2/M-phase microglia between day 2 and day 3 after nerve injury displayed distinct gene expression profiles. Analyses are two-way ANOVA with Sidak’s multiple-comparison test, n = 4–6, means ± SEM, *P < 0.05, ***P < 0.001, ****P < 0.0001, ns, not significant. The statistical analysis in b shows the difference between Myc cKO and control animals.