LYVE1^hi macrophages have their own gene expression patterns. (A) PCA of tissue-resident macrophages (alveolar macrophages, F4/80⁺ peritoneal macrophages, microglia, splenic red pulp macrophages, and LYVE1^hi membrane-associated macrophages) and blood monocytes (Ly6C^hi and Ly6C^lo monocytes) obtained from RNA-seq dataset. PC, principal component. (B) Heatmap analysis of top 50 up-regulated genes of 12,000 genes expressed in LYVE1^hi membrane-associated macrophages. Heatmap depicts mean expression intensity of mRNA transcripts for genes differentially expressed between LYVE1^hi macrophages and other macrophages, including monocytes. (C) Pathway analysis of genes differentially expressed in LYVE1^hi membrane-associated macrophages implemented by fast GSEA, showing top 10 enriched pathways from Reactome database and Molecular Signatures Database. NES, normalized enrichment score. (D) t-SNE plot displaying reanalyzed scRNA-seq of whole mesentery cells (accession no. GSE102665). (E) Expression of Lyve1, MMP9, and Folr2 on the t-SNE plot of scRNA-seq described in D. (F) Flow cytometric analysis showing FOLR2 expression of LYVE1^hi and LYVE1^lo mesenteric macrophages. Data are representative of three mice. (G) Violin plot of Retnla and Mrc1 expression obtained from scRNA-seq described in D.