Neutrophil Isolation from NOD-SCID Gamma Mice

Peripheral blood of NOD-SCID gamma mice was drawn from the subclavian vein after an intraperitoneal injection of a sublethal dose of anaesthesia (Avertin 0.2%, 0.16 ml/g [Sigma-Aldrich; St. Louis, MO, USA]) and collected in EDTA BD microtainer tubes (BD Biosciences; Franklin Lakes, NJ, USA). All mice were killed by cervical dislocation. After removing red blood cells using Quicklysis solution (Cytognos SL; Salamanca, Spain), neutrophils were isolated and purified following the Neutrophil Isolation Kit protocol (Miltenyi Biotec; Bergisch Gladbach, Germany [reference # 130-097-658])^{17 (link)–19} . The degree of the neutrophil purity achieved (CD11b + Ly6G+ cells; Table S3 in Supplementary Material) was assessed using standard sorting procedures by flow cytometry.