Fig. 2: Monocytes recruited to the IgE-CAI skin lesion sequentially differentiate into early and late CMDMs.

a–c Mo-Mac clusters shown in Fig. 1f were subjected to RNA velocity analysis. Stream plot of velocity vectors embedded in UMAP plot is shown. b, c Mo-Mac clusters from WT BALB/c mice shown in Fig. 1h were subjected to pseudotime trajectory analysis. UMAP plot colored by pseudotime is shown in (b). Gene expression changes of indicated genes along with pseudotime ordering are shown in (c). Colors indicate different clusters identified in Fig. 1h. d, e WT BALB/c mice were treated as in Fig. 1 to induce IgE-CAI. Cells isolated from the IgE-CAI skin lesion on days 0, 1, 3 and 5 were subjected to flow-cytometric analysis. In (d), the surface expression of PD-L2 and Ly6C in the monocyte-macrophage (Mo-Mac) populations is shown. In (e), time course of the number of indicated Mo-Mac subsets in the ear skin is shown (mean ± SEM, n = 3 biologically independent animals for Day 0–3, n = 4 for Day 4 and 5). f Ly6C^hi monocytes isolated from the bone marrow of CD45.1⁺ C57BL/6 mice were adoptively transferred to CD45.2⁺Ccr2^−/− C57BL/6 mice on day 1 post-challenge. Cells isolated from the IgE-CAI skin lesion on days 3 and 5 were subjected to flow-cytometric analysis. The surface expression of PD-L2 and Ly6C in the CD45.1⁺ Mo-Mac population is shown. g Ly6C^hi classical monocytes isolated from the bone marrow of WT or Il4ra^−/− BALB/c mice were incubated ex vivo with culture supernatants of bone marrow-derived basophils (BMBAs) that had been stimulated with anti-TNP IgE and TNP-OVA or control OVA for 18 h. The surface expression of Ly6C and PD-L2 in monocytes before the culture (left) and after 24 h- or 48 h-incubation with BMBA supernatants (middle and right, respectively) are shown. Data shown in (d, e, g) are representative of three independent experiments. Data shown in (f) are representative of two independent experiments. Source data are provided as a Source Data file.