Wild type c57bl 6 mice

Wild-type C57BL/6 mice were purchased from Samtako Bio Korea (Gyeonggi-do, Korea). Primary bone marrow–derived macrophages (BMDMs) were isolated from six-week-old C57BL/6 mice and cultured in DMEM for 3–5 days in the presence of M-CSF (R&D Systems, 416-ML), as described previously [32 (link)]. HEK293T (ATCC-11268; American Type Culture Collection) or THP-1 (ATCC-TIB-202) cells were maintained in DMEM or RPMI1640 (Gibco, NY, USA) containing 10% FBS (Gibco, NY, USA), sodium pyruvate, nonessential amino acids, penicillin G (100 IU/mL), and streptomycin (100 μg/mL). Transient transfections were performed using calcium phosphate (Clontech, Mountain View, CA, USA) in 293T, according to the manufacturer’s instructions. THP-1 stable cell lines were generated by transfections were performed using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA) and then a standard selection protocol with 400–800 μg/mL of G418.

Wild‐type C57BL/6 mice were purchased from Samtako Bio Korea (Gyeonggi‐do, Korea). Primary bone marrow‐derived macrophages (BMDMs) were isolated from C57BL/6 mice and cultured in DMEM for 3–5 days in the presence of M‐CSF (R&D Systems, 416‐ML), as described previously (Koh et al, 2017). BMDMs of TLR2^−/−, TLR4^−/−, MyD88^−/−, TRIF^−/−, IRAK1^−/−, TRAF6^−/−, and TBK1^−/− in C57BL/6 mice were a generous gift from Dr. Chul‐Ho Lee (Laboratory Animal Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea). HEK293T (ATCC‐11268; American Type Culture Collection) and RAW264.7 (ATCC TIB‐71) in DMEM (Gibco) containing 10% FBS (Gibco), sodium pyruvate, nonessential amino acids, penicillin G (100 IU/ml), and streptomycin (100 μg/ml). Human monocytic THP‐1 (ATCC TIB‐202) cells were grown in RPMI 1640/glutamax supplemented with 10% FBS and treated with 20 nM PMA (Sigma‐Aldrich) for 24 h to induce their differentiation into macrophage‐like cells, followed by washing three times with PBS. Transient transfections were performed using calcium phosphate (Clontech) in 293T, according to the manufacturer’s instructions. RAW264.7 and THP‐1 stable cell lines were generated by transfections were performed using Lipofectamine 3000 (Invitrogen) and then a standard selection protocol with 400–800 μg/ml of G418.

Wild-type C57BL/6 mice were purchased from Samtako Bio (Osan, Gyeonggi-do, Korea) and maintained under specific pathogen-free conditions in the animal facilities at Chungnam National University School of Medicine. Mice were used in accordance with the guidelines of the Institutional Animal Care and Use Committee, Chungnam National University School of Medicine, Daejeon, Korea (approval nos. 202109A-CNU-180 and CNUH-021-A0011).