Penicillin streptomycin

MDCC-MSB1 cells (a chicken lymphoma cell line) cells were kept in our lab. DF-1 cells (a chicken embryo fibroblast cell line) cells were purchased from American Type Culture Collection (ATCC). MDCC-MSB1 cells were cultured in Roswell Park Memorial Institute 1640 medium (Invitrogen, Carlsbad, the United States) supplemented with 10% fetal bovine serum (FBS) (Gibco, Grand Island, the United States) and 1% penicillin-streptomycin (MACGENE Technology). DF-1 cells were cultured in Dulbecco Modified Eagle Medium (DMEM) (Invitrogen, Carlsbad, the United States) supplemented with 10% FBS (Gibco, Grand Island, the United States) and 1% penicillin-streptomycin (MACGENE Technology). All cells were maintained at 37 °C in a humidified atmosphere incubator with 5% CO₂.
CIAV Cux-1 strain (GenBank accession number M55918.1) was kindly provided by Professor Bing Yang (Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agricultural and Forestry Sciences).

Using WT or Nlrp3^−/− female C57BL/6 mice (10-week-old), bone marrow cells were extracted into Dulbecco's modified Eagle's medium (DMEM, Macgene, Beijing, China) containing 1% penicillin/streptomycin (Macgene) and 10% fetal bovine serum (Gibco, Rockford, IL, USA). Moreover, 50 ng/mL murine macrophage colony-stimulating factor (MedChemExpress, Monmouth, NJ, USA) was added to DMEM to differentiate cells into bone marrow-derived macrophages (BMDMs). Human THP-1 cells, which were supplied by Dr. Tao Li from NCBA, were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (Macgene) containing 1% penicillin/streptomycin (Macgene) and 10% fetal bovine serum (FBS), and they were stimulated using 100 nmol/L phorbol-12-myristate-13-acetate (PMA, MedChemExpress) overnight to differentiate into macrophages. All of the cultured cells were placed in a humidified 5% CO₂ atmosphere at 37 °C.