Fig. 3: Late CMDMs display higher efferocytic ability than do early CMDMs. | Nature Communications

Fig. 3: Late CMDMs display higher efferocytic ability than do early CMDMs.

From: Single-cell transcriptomics identifies the differentiation trajectory from inflammatory monocytes to pro-resolving macrophages in a mouse skin allergy model

Fig. 3

Early and late CMDMs were compared by using GSEA. Enrichment plot of genes involved in lipid catabolic process (a) and phagocytosis (c). b WT BALB/c mice were treated as in Fig. 1 to induce IgE-CAI. Classical monocytes, early CMDMs and late CMDMs isolated from TNP-OVA-treated ear skins, and resident macrophages isolated from OVA-treated ear skins were subjected to Diff-Quik staining. Bars indicate 10 μm. d Dot plot showing the expression level of indicated genes in classical monocytes, early CMDMs, late CMDMs and resident-like macrophages. e–h WT BALB/c mice were treated as in Fig. 1 to induce IgE-CAI. Apoptotic neutrophils (e, f) or OVA (g, h) labeled (open histograms) or unlabeled (shaded histograms) with AcidiFluor were intradermally administered to the ear skin on day 5 post-challenge. Two hours after the injection, skin samples were subjected to flow cytometric analysis. In (e) and (g), histograms of AcidiFluor fluorescence in Mo-Mac subsets are shown. In (f) and (h), the mean fluorescence intensity (MFI) of AcidiFluor in Mo-Mac subsets is shown (mean ± SEM, n = 3 biologically independent animals each). In (f), **p = 0.0096 (mono vs. early CMDM); ***p = 0.0001 (mono vs. late CMDM) *p = 0.0144 (early vs. late CMDM), ***p = 0.0002 (late CMDM vs. resident-like) measured by one-way ANOVA with Tukey’s multiple comparison test. In (h), *p = 0.0135 (mono vs. early CMDM); ****p = 3.36 × 10−5 (mono vs. late CMDM) **p = 0.0014 (early vs. late CMDM), *p = 0.0218 (late CMDM vs. resident-like) measured by one-way ANOVA with Tukey’s multiple comparison test. i WT and Ccr2−/− BALB/c mice were treated as in (e) and (g). Histograms of AcidiFluor fluorescence in total Mo-Macs are shown in left panels. The MFI of AcidiFluor in Mo-Mac populations is shown in right panels (mean ± SEM, n = 6 and n = 4 biologically independent animals for apoptotic neutrophil uptake and OVA uptake, respectively). **p = 0.0082 (for apoptotic neutrophil) *p = 0.0196 (for OVA) measured by two-sided unpaired Student’s t test. Data shown in (b, e–i) are representative of three independent experiments. Source data are provided as a Source Data file.

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