Fig. 2: The distinction between Flt3 and Tie2 in Ly6C⁺ and Ly6C⁻ AMs.

Gating strategy for AMs (CD45 ⁺ Siglec-F ⁺ CD11b-CD11c ⁺ CD64 ⁺ ), CD11b ⁺ DCs (CD11b ⁺ MHC-II ⁺ CD11c ⁺ CD24 ⁺ CD64-), CD103 ⁺ DCs (CD11c ⁺ CD103 ⁺ CD24 ⁺ ), eosinophils (Siglec-F ⁺ CD11b ⁺ CD11c-), interstitial macrophages (CD11b ⁺ MHC-II ⁺ CD11c ⁺ CD64 ⁺ CD24-), Ly6C⁺ monocytes and macrophages, Ly6C⁻ monocytes and macrophages (CD11b ⁺ MHC-II-CD64⁺/−), and neutrophils (CD11b⁺ Ly6G⁺) in the mouse lung and the RGS1 expression in the cells were evaluated (a). Gating strategy for endovascular progenitor (EVP, CD45-CD31^lowVEGFR^low), transit-amplifying (TA, CD45-CD31^intVEGFR^int), and definitive differentiated cells (D, CD45-CD31^hiVEGFR^hi) and their RGS1 expression were evaluated (b). The expression of Flt3 and Tie2 was evaluated in Ly6C⁺ or Ly6C⁻ brain macrophages (c), intestinal macrophages (d), and lung macrophages (e). Construction mode diagrams of RGS1^fl/fl;Flt3-CreERt2 and RGS1^fl/fl;Tie2-CreERt2 (f). One week after tamoxifen treatment, the expression of RGS1 in RGS1^Flt3 and RGS1^Tie2 mice in Flt3 ⁺ Ly6C^hi AMs, Flt3 ⁺ Ly6C^low AMs, and Tie2 ⁺ Ly6C⁻ AMs was detected (g, h). The data are shown as the mean ± SD; *p < 0.05 and **p < 0.01 by ANOVA, and “n.s.” indicates not statistically significant. The data in a-e were derived from 3 independent experiments. The data in g and h are based on 3 mice in each group.