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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. 10 — a Quantification of Sox9⁺C3⁺ astrocytes in the spinal cord of IL-1R1-deficient (Il1r1^r/r) and cell-specific IL-1R1 conditional restored mice injected i.c.m. with PBS or rmIL-1α and killed at day 1 (n = 15 WT + PBS, n = 16 WT + rmIL-1α, n = 13 Il1r1^r/r + PBS, n = 11 Il1r1^r/r + rmIL-1α, n = 3 Pdgfra^CreER::Il1r1^r/r + PBS, n = 5 Pdgfra^CreER::Il1r1^r/r + rmIL-1α, n = 2 Cx3cr1^CreER::Il1r1^r/r + PBS, n = 3 Cx3cr1^CreER::Il1r1^r/r + rmIL-1α, n = 3 Cdh5^CreER::Il1r1^r/r + PBS, n = 6 Cdh5^CreER::Il1r1^r/r + rmIL-1α, n = 3 Gfap^Cre::Il1r1^r/r + PBS, n = 4 Gfap^Cre::Il1r1^r/r + rmIL-1α). b, c Sox9 (red) and C3 (green) immunostainings in the spinal cord of mice injected with PBS or rmIL-1α. White arrowheads point to double-labeled cells. d Experimental design for the lactate dehydrogenase (LDH) assay. Primary astrocytes were cultured in presence of PBS (vehicle) or rmIL-1α. Primary mature OLs were then incubated in DMEM containing (or not) rmIL-1α, or conditioned medium derived from astrocytes (ACM) stimulated with vehicle or IL-1α. e Quantification of OL loss using LDH assay (n = 3 wells/condition). f Quantification of reactive oxygen species (ROS) production in primary astrocytes stimulated with vehicle or rmIL-1α (n = 6 wells/condition). g, h Immunofluorescence showing Ly6G⁺ neutrophils (red) in the spinal cord of C57BL/6 mice injected i.c.m. with rmIL-1α and i.p. with N-acetyl-L-cysteine (NAC) or saline. Mice were killed at 24 h post-i.c.m. injection. i Quantification of spinal cord-infiltrated Ly6G⁺ neutrophils (n = 8 Saline+PBS, n = 9 Saline+rmIL-1α, n = 8 NAC + PBS, n = 7 NAC + rmIL-1α). j, k Confocal immunofluorescence showing Olig2 (red) and CC1 (green) in the spinal cord of mice injected i.c.m. with rmIL-1α and i.p. with NAC or saline at 24 h. l Quantification of mature OLs in the spinal cord (n = 8 mice/group). m, n Immunostaining of Ly6G⁺ neutrophils (red) at the lesion epicenter at day 1 post-SCI in C57BL/6 mice treated with NAC or saline. o Quantification of spinal cord-infiltrated neutrophils (n = 5 mice/group). p, q Immunostaining for Olig2 (red) and CC1 (green) at the lesion epicenter at day 1 post-SCI. r Quantification of mature OLs in the injured spinal cord (n = 5 mice/group). Data are means (+/−SEM) and statistical significance determined by two-way ANOVA followed by Bonferroni post-hoc test (a, e, f, i, l, o, r). **p < 0.01 (r). Other pairwise comparisons and p-values are indicated in graphs. All scale bars: 50 µm.