LNs have a distinct immunophenotype and mRNA expression profile compared with other neutrophils. (A) Neutrophil proportions from total bone marrow, peripheral blood, lung, and spleen (n = 3) from 8-week-old C57BL/6 male mice were analyzed by flow cytometry. (B) Heatmap of Ly6G expression on bidimensional UMAP analysis of live CD45⁺ immune cells from bone marrow, peripheral blood, lung, and spleen (n = 3). (C) Three Ly6G^high clusters among 38 clusters from PARC analysis in bone marrow, blood, lung, and spleen. (D) Cluster proportion of each tissue neutrophil. Expression levels (MFI) of CD11b, CD62L, CXCR4, MHC II, and CD101 on Ly6G⁺ clusters (E) and tissue neutrophils (F). Correlation matrix (G) and up/down (log₂FC ≥ 1/log₂FC ≤ 1) (H) regulated count of genes from bulk RNA-seq data of BMNs, BNs, and LNs with log₂FC ≥ 1, P < .05 from pooling 5 heads of 8-week-old C57BL/6 male mice. (I) Gene set enrichment analysis (GSEA) data of gene sets associated with translational activity in LNs compared with BNs. (J) Heatmap of surface protein mRNA expression on BMN, BN, and LN in RNA-seq. (K) Surface protein level (MFI) of Sca1, Plet1, and CD14 on tissue neutrophils (n = 4). Data are shown as mean ± standard error of the mean. Significant differences are denoted as *P < .05; **P < .01; ***P < .001; ****P < .0001. ns, not significant (one-way ANOVA) (A [right], E-F,K).