*4.8. Isolation of Peritoneal Neutrophils*

Thioglycolate-elicited peritoneal neutrophils were obtained from WT mice by a previously described method [39]. WT mice were intraperitoneally injected with 1.5 mL of sterile 4% thioglycolate; 12–15 h later, peritoneal lavages were performed with 10 mL PBS in each mouse. The proportion of neutrophils in the lavages was over 90% as assessed by FACS analysis of anti-CD45, anti-CD11b, and anti-Ly6G expression. Peritoneal neutrophils were recovered by centrifugation and suspended in RPMI 1640 medium supplemented with 10% FCS, 100 U/mL penicillin G, 100 μg/mL streptomycin, and 50 μM 2-mercaptoethanol. The obtained cells were cultured at 1 × 10<sup>6</sup>/mL with various doses of IFN-γ or lipopolysaccharide (LPS) (Sigma-Aldrich, St. Louis, MO, USA) for 24 h at 37 ◦C.
