PMNs were isolated and cultured as described previously (21 (link)). In brief, tibias and femurs harvested from WT mice and bone marrow was flushed with Dulbecco’s modified Eagle’s medium (DMEM) (22 ). Lung PMN were collected from bronchoalveolar lavage (BAL) fluid. For BAL (23 (link)), mice were euthanized by an overdose of Pentobarbital. Lungs were lavaged with 0.5 mL sterile saline each time (up to 10 times) using an intratracheal catheter. Bone marrow cell pellets and BAL cells were collected by centrifugation at 4°C, and erythrocytes were lysed with RBC lysis buffer (eBioscience, San Diego, Calif). The immunomagnetic separation system (BD Biosciences, San Jose, Calif) was used to isolate PMN from bone marrow and BAL fluid (24 (link)). The anti-mouse Ly6G and Ly6C particle magnetic nanoparticle-conjugated antibodies were chosen to label and isolate PMNs. The purity and viability of the isolated PMNs are >94% and >90%, respectively.