Extended Data Fig. 4: Validation of antibody-mediated depletion of various cell populations.

(a-e) The efficiency of antibody-mediated depletion of NK cells, neutrophils, and the targeting of CCR2⁺ monocytes was determined by flow cytometry 2 days after antibody administration. The abundance of the depleted population in peripheral blood (for NK (n = 3), neutrophils (n = 3) and CCR2⁺ monocytes (n = 5)) or spleen (for CCR2⁺ monocytes (n = 5)) was calculated out of total CD45⁺ leucocytes and compared to matched isotype control by two-tailed unpaired t-test (one independent experiment). NK cells were defined as CD45⁺/CD11b⁻/CD3⁻/B220⁻/NK1.1⁺. Neutrophils were defined as CD45⁺/CD3⁻/CD11b⁺/SSC^high/Gr1⁺ (see Extended Data Fig. 2b for gating strategy). CCR2⁺ monocytes were defined as CD45⁺/CD19⁻/CD3⁻/NK1.1⁻/CD11b⁺/Ly6G⁻/Ly6C⁺. (f-i) Ifnar1^−/−/FcγR humanized mice were treated with clodronate liposomes or control liposomes to deplete macrophages. Depletion of macrophages in the spleen and liver as well as efficiency of depletion on multiple myeloid subsets in the spleen (macrophages, neutrophils, and monocytes) was determined 2 days post treatment by flow cytometry. Macrophages (Mac) were defined as CD45⁺/CD19⁻/CD3⁻/SiglecF⁻/Gr1⁻/CD11b⁺/F4/80⁺. Red pulp macrophages (RPM) were defined as CD45⁺/CD19⁻/CD3⁻/SiglecF⁻/Gr1⁻/CD11b^low/F4/80⁺. Neutrophils were defined as CD19⁻/CD3⁻/SiglecF⁻/SSC^high/Gr1^high. Monocytes were defined as CD19⁻/CD3⁻/SiglecF⁻/SSC^low/Gr1^intermediate. Groups were compared by two-tailed unpaired t-test. n = 3 mice per group in one experiment. (j) To determine whether splenectomy affect platelet numbers in circulation, platelet counts were determined pre- and post- splenectomy and compared by two-tailed unpaired t-test (n = 5 mice per group in one experiment).

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