Lack of D6 on the nonhematopoietic compartments causes increased number of inflammatory monocytes. (A) Percentage in CD45⁺ gate (left panel) and fold increase of absolute number (right panel) of CD11b⁺/Ly6C^high monocytes recovered from blood in D6^−/− (black bar) versus WT mice (white bar). (B) Fold increase of percentage of CD11b⁺/Ly6C^high monocytes in total BM CD45⁺ leukocytes of D6^−/− (black bar) and WT mice (white bar). (C) Gating strategy for evaluating the percentage and absolute numbers of leukocytes in mouse spleen. Dead cells, doublets, and CD3⁺, CD19⁺, and NK1.1⁺ cells were excluded for further analysis. Cell labeling with anti-CD11b and CD11c allows identification of myeloid DCs (R1: CD11b⁺/CD11c^high), red pulp macrophages (R2: F4/80⁺), and plasmacytoid DCs (R3: PDCA-1⁺) in the CD11c^dim/CD11b^dim gate. Inflammatory monocytes (R4) and neutrophils (R5) are distinguished within the CD11b^high population as SSC^low/CD115⁺/Ly6C^high and SSC^high/CD115⁻/Ly6G^high, respectively. Gates are based on isotype controls. (D) Fold increase of the absolute number of cells in spleen of D6^−/− (black bar) versus WT (white bar) mice based on gated strategy used in panel C. (E) Fold increase of the absolute number of CD11b⁺/Ly6C^high monocytes in blood of D6^−/− (Ly5.2) mice reconstituted with WT Ly5.1 or D6^−/− BM cells and in WT mice (Ly5.1) reconstituted with D6^−/− BM cells versus WT mice (Ly5.1) reconstituted with WT mice (Ly5.2). In all panels, data are representative of 4 independent experiments (n = 10 mice/experiment). *P < .05. **P < .005.