Extended Data Fig. 4: Neutrophil subsets in the blood, brain and BM of young and aged stroke mice. | Nature Immunology

Extended Data Fig. 4: Neutrophil subsets in the blood, brain and BM of young and aged stroke mice.

From: Age-induced alterations of granulopoiesis generate atypical neutrophils that aggravate stroke pathology

Extended Data Fig. 4: Neutrophil subsets in the blood, brain and BM of young and aged stroke mice.The alternative text for this image may have been generated using AI.

a, Quantification of circulating neutrophil subsets by 18-color MCFC at 0h, 6h and 24h poststroke, (n = 4 to 6 mice/group/ timepoint, 2-ANOVA, Bonferroni-post-hoc test, **p ≤ 0.01; ****p≤0.0001). b, Heatmap displaying the median antigen intensity in blood neutrophil clusters analyzed by MCFC at 24h poststroke and dotplot showing the scaled expression of selected genes, corresponding to markers of MCFC, of the scRNAseq stroke blood neutrophil subsets at 24h poststroke (neutrophil subsets from Fig. 3). The table shows the nomenclature and marker antigens and genes for major neutrophil subsets and GSVA based comparison (shown in Fig. 3g) with the nomenclature used by Xie et al.17. c, Histograms, and barplots showing the median fluorescence intensity (MFI) of CD177, CD11b and CD101 on neutrophils infiltrating the ischemic lesion analyzed by MCFC at 24h poststroke in young and aged mice (n = 5 mice/group, 2-tailed, Wilcoxon Mann-Whitney, *p = 0.031, **p = 0.0079, each dot corresponds to a single mouse). d, Representative IF images of CD177, CD101 or CD14 (in red) on neutrophils (Ly6G+in green, or white and 7/4+ in white) infiltrating the ischemic lesion of aged mice at 24h poststroke (blood vessels, stained by t-lectin, white) Scale bar, 25µm. e, Quantification of neutrophils and progenitors in the BM by MCFC at 0h and 24h poststroke of young and aged mice (n = 4- 6 mice per group per timepoint, 2-ANOVA Bonferroni post-hoc test; *p ≤ 0.05; **p ≤ 0.01; ****p≤0.0001). f, Barplots showing frequencies among mature BM neutrophils or BM neutrophil at 6h poststroke (n = 3 young and 4 aged mice, 2-tailed t-test, *p ≤ 0.05). g, ELISA for selected blood cytokines and chemokines at 0h and 48h poststroke; (n = 5 to 7 mice per group per timepoint, 2-ANOVA Bonferroni post-hoc test *p ≤ 0.05; **p≤0.01, ***p ≤ 0.001; §p≤0.055). In a, c, e and g, the box extends from the 25th- 75th percentiles, midline at median, whiskers plot min and max.

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