Fig. 4: Heterogeneous cellular composition and phenotypic characteristics in neutrophils.

A The t-SNE map of neutrophils shows the expression of Ly6G. B The t-SNE map shows the cluster partitions. C t-SNE plots of neutrophils derived from the brain samples of five groups are shown. Colors indicate different groups. D, E A heatmap displays the median marker expression value and hierarchical clustering of the markers for 9 clusters identified in B. Vertical bar graph depicting the composition of neutrophils in mice brain in indicated time points. The colored segment lengths represent the proportion of cells as a percentage of total neutrophils in each sample. Colors as in D. F t-SNE maps show the expression of PD-L1 and SS2 in neutrophils. G Apoptosis of PD-L1⁺ neutrophils and PD-L1^- neutrophils were detected by flow cytometry. Four female mouse brains were pooled in one sample, N = 3. H The phagocytic capacity of PD-L1⁺ neutrophils and PD-L1^- neutrophils was detected by flow cytometry. Four female mouse brains were pooled in one sample, N = 3. I Laser confocal results showed the expression of H3 and MPO in PD-L1⁺ PMN and their negative counterpart. Scale bar = 10 μm. J Median expression and statistical plots of CD14 and TLR2 expression on neutrophils in indicated groups. N = 4. Error bars represent the median ± SEM. A one-way ANOVA or Dunnett’s test was performed for analysis of multiple groups. Student’s t-test was used to compare the two groups. P value < 0.05 (*); p < 0.01 (**); p < 0.001 (***); NS no significant difference.