Anti cd64 clone x54 5 7

Blood was prepared by ACK lysis. Single cell suspensions of brain were prepared as previously described (17 (link), 38 (link)). Mice were euthanized with CO₂ and transcardially perfused with ice-cold HBSS. Brains were grossly free of blood. Whole brain was minced, and either mechanically triturated (in experiments for determination of relative proportions of infiltrating cell types) or enzymatically homogenized (in cell sorting experiments) with Neural Tissue Dissociation Kit with Papain, Miltenyi) and passed through a 70 μm cell strainer to form a single cell suspension. Cells were enriched by centrifugation over a discontinuous gradient of HBSS, 37% and 70% Percoll and collecting the fraction at the 37%/70% boundary. Cells were then washed and stained with fluorophore conjugated antibodies prior to analysis on a FACSAria II flow cytometer and cell sorter (BD). Antibodies include anti-CD11b (clone M1/70, BD), anti-CD45 (clone 30-F11, BD), anti-CD64 (clone X54-5/7.1, BD), anti-Ly6G (clone 1A8, Biolegend), anti-Ly6C (clone HK1.4, Biolegend). Propidium iodide was used as a live/dead marker. Gating was performed as described in Figure S1.