Bone Marrow–Derived Macrophage Preparation

The femur and tibia of WT C57BL/6 mice were dissected. Bone marrow was flushed with 10 mL of RPMI-1640 medium and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and cell pellets were treated with eBioscience1× RBC Lysis Buffer. After washing, the cells were cultured in RPMI-1640 medium supplemented with 10 % fetal bovine serum and mouse macrophage colony-stimulating factor (Thermo Fisher). The medium was replenished every 3 days and day-6 macrophages were used for the experiment.