Nicotinamide adenine dinucleotide

MG strain (R_low) was purchased from China Veterinary Culture Collection Center. MG were grown at 37 °C in 5% CO₂ in Mycoplasma basal medium (BasalMedia, Shanghai, China) supplemented with 20% fetal bovine serum (Gibco Hyclone, Shanghai, China), 0.1% Nicotinamide adenine dinucleotide (Solarbio, Beijing, China) and 0.05% penicillins (Beyotime, Shanghai, China). MG were prepared for subculture every 3 to 5 d, the passage 3 used in the subsequent experiments.

Poly-inosinic-polycytidylic acid (Poly IC) was purchased from Sigma-Aldrich (St. Louis, MO). CCK-8 cell proliferation and cytotoxicity assay kit, nicotinamide adenine dinucleotide (NAD), RBC Lysing Buffer, and other reagents were obtained from Solarbio. Tumor necrosis factor (TNF)-α, interferon (IFN)-β, interleukin (IL)-1β, IL-23, IL-17, and IL-22 ELISA kits were purchased from Neobioscience. Bradford Protein Assay Kit was obtained from Beyotime Biotechnology. For flow cytometry, Intracellular Fixation/Permeabilization Buffer Kit, purified anti-mouse CD16/32, FITC-CD11c, PE-MHC II, APC-CD80, PerCP-CD86, PE-F4/80, APC-CD11c, and PerCP-Ly6G were from Elabscience; FITC-lineage cocktail and APC-RORγt were from eBioscience. B5 (amino acid sequence: NPQSCRWNMGVCIPISCPGNMRQIGTCFGPRVPCCRRW) was synthesized at Shanghai Apeptide Co., Ltd. M.W.: 4325.12; Purity: >95%; B5 is soluble in water, grand average of hydropathy: −0.268; Isoelectric point: 8.91; B5 stock solution (2 mg/ml in PBS) for experiments. PBD2 (amino acid sequence: DHYICAKKGGTCNFSPCPLFNRIEGTCYSGKAKCCIR) was synthesized at Shanghai Apeptide Co., Ltd. Purity: >95%.