GM332_2
SAMPLE8 Bone marrow was flushed from femurs and tibias of B6 mice. BMDMs were harvested in complete DMEM medium (DMEM supplemented with 1% penicillin/streptomycin and 5% FBS) containing GM-CSF (50 ng/mL) for 7 days. On day 3 and day 6, non-adherent cells were discarded, and then fresh GM-CSF-containing complete DMEM medium was added. For macrophage subset screening, a total of 22 cytokines or stimuli were used to treat sorted PEMs for 48 h in complete DMEM medium. For the M1 subset induction, macrophages were treated with 100 ng/mL LPS for 6 h, the M2 subset was induced by adding IL-4 (10 ng/mL) for 48 h, and the M17 subset were obtained by treatment with IL-23 (10 ng/mL) for 48 h 20. We defined 75 ng/mL IL-33-treated macrophages as the M(IL-33) subset and the M(IL-33+IL-2) subset were induced by IL-33 (75 ng/mL) and IL-2 (10 ng/mL) for 48 h. The untreated macrophage was M0.