Induction of BMDMs

L929-conditioned medium was obtained from L929 cells plated in a 10-cm dish containing 10 mL of L929 medium for 3 days to generate macrophage colony-stimulating factor. Primary bone marrow-derived macrophages (BMDMs) were obtained from bone marrow cells (BMCs), which were flushed from the bones of BALB/c mice and filtered through nylon mesh. BMCs were differentiated into macrophages in BMDM medium containing complete RPMI1640 medium, 20% L929-conditioned medium, 20% FBS, and 100 U/mL penicillin, and 100 μg/mL streptomycin. Cells were cultured in a 10 cm dish at a density of 3 × 10⁵ cells/mL and maintained in 5% CO2, 37°C incubator. Six days after initial BMC cell culture, cell surface antigen F4/80 was detected by flow cytometry, the cellular differentiation reached a purity of about 90%. Stable overexpressing BMDMs-AIM2 cells were obtained by infection with pCDH—CMV-MCS-EF1-AIM2 lentivirus generated by HEK293T. The same amount of vector lentivirus was used as a control.