SPF C57BL/6 mice (Female, 5-week-old, 16–18g) were purchased from Anhui Laboratory Animal Center (Hefei, China). All mice were housed at a temperature of 20–26°C and on a 12 h light and dark alternating time. This study was conducted in accordance with the Basel Declaration and the ethical guidelines by the International Council for Laboratory Animal Science (ICLAS). The Animal Care and Use Committee of Anhui Medical University approved all experimental procedures (Permit Number: LLSC20170361).
The murine macrophage cell line RAW264.7 was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Sigma Inc., St. Louis, MO), supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY) and 1% penicillin/streptomycin solution (C0222, Beyotime Biotechnology, China) at 37°C in 5% CO2. RAW264.7 cells were grown in 24-well plates to 70–80% confluence and then stimulated with OVA (500 μg/mL, Sigma). Some cells were treated with CpG-ODN 1668 (1.5 μM, 5 μM, Sangon, Shanghai, China) for 1 h prior to stimulation with OVA. Some cells were treated with SP600125 (10 μM, Sigma Inc., St. Louis, MO) for 2 h prior to stimulation with OVA.
The murine macrophage cell line RAW264.7 was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Sigma Inc., St. Louis, MO), supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY) and 1% penicillin/streptomycin solution (C0222, Beyotime Biotechnology, China) at 37°C in 5% CO2. RAW264.7 cells were grown in 24-well plates to 70–80% confluence and then stimulated with OVA (500 μg/mL, Sigma). Some cells were treated with CpG-ODN 1668 (1.5 μM, 5 μM, Sangon, Shanghai, China) for 1 h prior to stimulation with OVA. Some cells were treated with SP600125 (10 μM, Sigma Inc., St. Louis, MO) for 2 h prior to stimulation with OVA.