Additional ontogeny data and response to peripheral inflammation by the two macrophage subsets. Additional analysis of DRG mural cells. (A) Mouse fur at 27 wk after hindleg irradiation. (B) Flow cytometry–based analysis of hindleg-irradiated CD45.1:Cx3cr1^GFP/+→ CD45.2 chimeric mice at 33 wk after irradiation. Chimerism was calculated as frequency of GFP⁺ cells within indicated cell subsets, normalized to blood GFP⁺ frequency. n = 5 mice. The experiment was performed once. Student’s unpaired t test. ***P < 0.001. (C) Expression of TLF genes (Dick et al., 2022) across monocyte/macrophage/DC clusters in scRNA-seq data from Fig. 4 A. (D) Expression of TLF markers by flow cytometry in indicated cell types using flow cytometry. n = 4 mice pooled. The experiment was performed twice. (E) FOLR2 expression in indicated macrophage subsets. (F) Experiment illustration for macrophage depletion and peripheral LPS challenge. Mice were fed control or PLX3397 chow (290 ppm) and injected i.p with 1 mg/kg LPS or saline. Created with https://BioRender.com. (G) Quantification of ICAM1 expression in CD31⁺ vessel segments. Two-way ANOVA with Tukey’s multiple comparisons test. n = 5 mice/group. The experiment was performed once. *P < 0.05, ***P < 0.001. Scale bar, 20 μm. A, artery; C, capillary; V, vein. (H) Quantification of endothelial coverage by Iba1⁺ perivascular macrophages, split into CD163⁺ and CD163⁻ subsets. Endothelial coverage was additionally analyzed based on which vessel segment macrophages contacted. n = 5 mice/group. The experiment was performed once. Sidak’s multiple comparisons tests. *P < 0.05, ***P < 0.001; ns, not significant. (I) mRNA expression of indicated fibroblast, SMC, and pericyte marker genes in the dataset used for CellChat (Avraham et al., 2020), presented in Fig. 7, A and B. (J)Pdgfrb^GFP/+ mice label GFP⁺ACTA2⁺ SMCs covering arteries and veins. Scale bar, 50 μm.