The cells were washed in FACS buffer (1% BSA, 0.01% sodium azide, 2.5 mM EDTA in sterile PBS; 650 g, 10 min, 4°C) and preincubated for 15 min with anti-CD16/CD32 Fc-block antibody (10 μg/mL; BioLegend, San Diego, CA, 1:50) in FACS buffer followed by a 30 min incubation with fluorescence dye-conjugated, anti-mouse antibodies for SiglecF (clone E50–2440, 1:400), CD11c (clone N418, 1:100), CD11b (clone M1/70, 1:100), Ly6G (clone 1A8, 1:50, clone 1A8, 1:400), Ly6C (clone HK1.4, 1:80/1:600), F4/80 (clone BM8, 1:100), CD62L (clone DREG-56, 1:80), CD66b (clone G10F5, 1:40), and corresponding isotype controls (all from BioLegend). Analysis was done in adherence to guidelines [25 (link)].
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