Mouse femurs and tibias were flushed with macrophage culture medium [RPMI 1640 containing 100 U/ml of penicillin/streptomycin, 1 mM Hepes (pH 7.4), and 10% fetal bovine serum (FBS)], and the bone marrow was filtered through a 70-μm cell strainer. Cells were thereafter incubated in macrophage culture medium supplemented with macrophage colony-stimulating factor (M-CSF) (100 ng/ml) for 6 days. BMDMs were then reseeded and polarized overnight toward M1 or M2 with IFN-γ (50 ng/ml) plus LPS (20 ng/ml) or with IL-4 (20 ng/ml), respectively.

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