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. 2022 Oct 2;13:5786. doi: 10.1038/s41467-022-33463-x

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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. 9 — a Diagram showing the experimental design and timeline of the depletion study. Abbreviations: Combo, combination strategy; Combo⁺, adapted combination strategy; i.c.m., intra-cisterna magna; i.p., intraperitoneal; sac, sacrifice. b Quantification by flow cytometry of the number of Ly6G^TdT+ neutrophils in the blood of PBS- and IL-1α-injected Ly6g^Cre-TdT::R26-TdT mice pretreated with either the Combo⁺ strategy, Combo strategy, isotype control antibodies or saline. Mice were killed at 24 h post-i.c.m. injection (n = 3 Saline + PBS, n = 5 Isotype control + PBS, n = 4 Combo + PBS, n = 3 Combo⁺ + PBS, n = 3 Saline + IL-1α, n = 7 Isotype control + IL-1α, n = 4 Combo + IL-1α, n = 3 Combo⁺ + IL-1α). c Quantification of Ly6G^TdT+ neutrophils that infiltrated the spinal cord at 24 h post-i.c.m. injection (n = 3 Saline + PBS, n = 6 Isotype control + PBS, n = 4 Combo + PBS, n = 3 Combo⁺ + PBS, n = 4 Saline + IL-1α, n = 7 Isotype control + IL-1α, n = 4 Combo + IL-1α, n = 3 Combo⁺ + IL-1α). d, e Confocal images showing Ly6G^TdT+ cells (red) and laminin (green) in the spinal cord of Ly6g^Cre-TdT::R26-TdT mice injected i.c.m. with rmIL-1α, and pretreated i.p. with either isotype control antibodies (d) or the Combo⁺ treatment (e). f, g Confocal immunofluorescence imaging of Olig2 and CC1 in the spinal cord white matter of Ly6g^Cre-TdT::R26-TdT mice injected i.c.m. with either IL-1α or PBS, and treated with either isotype control antibodies (f) or the Combo⁺ strategy (g). h Quantification of Olig2⁺ CC1⁺ mature OLs in the spinal cord white matter of mice (n = 3 Saline + PBS, n = 6 Isotype control + PBS, n = 4 Combo + PBS, n = 3 Combo⁺ + PBS, n = 4 Saline + IL-1α, n = 7 Isotype control + IL-1α, n = 4 Combo + IL-1α, n = 3 Combo⁺ + IL-1α). i Percentage and total number (in parentheses) of Ly6G^TdT+ neutrophils in the spinal cord gray matter (GM) versus white matter (WM). All data are mean values + /− SEM and statistical significance was determined by a two-way ANOVA followed by Bonferroni post-hoc test (b, c, h). Pairwise comparisons and p-values are indicated in graphs. Scale bars: (d, e, in e) 50 µm, (f, g, in g) 50 µm.