iv) Wild-type vs XID mice in 16 h ZIP model
Male CBA/CaCrl (WT) or XID mice (8-10 weeks old) were given zymosan challenge (100 ug; i.p.). Blood was collected by direct cardiac puncture, peritoneal exudate cells were harvested in ice-cold PEC harvest buffer (PBS, 5 mM EDTA) and spleen dissected 16 h after zymosan challenge.
Murine bone marrow‐derived macrophages (BMDMs):
Bone marrow‐derived macrophages were generated as previously described (Recio et al., 2018). Briefly, fresh bone marrow cells from tibiae and femurs of male C57BL/6 or XID mice aged 8–10 weeks were cultured in DMEM containing 4.5 g·L−1 glucose, 2 mM l‐glutamine, 50 units·ml−1 penicillin and 50 μg·ml−1 streptomycin, 10% heat‐inactivated FBS, 10% L929 cell‐conditioned media (as a source of macrophage colony‐stimulating factor) and for 7 days. Bone marrow cells were seeded into 8 ml of medium in 90 mm non‐tissue culture treated Petri dishes (ThermoFisher Scientific, Sterilin, UK). On Day 5, an additional 5 ml of medium was added. Gentle scraping was used to lift cells. BMDMs were then counted and suspended in FBS free media at the desired cell concentration.