Batf3 ko

WT C57BL/6 and BALB/c mice were purchased from Charles River Laboratories. Rag1^KO, Ifng^KO, Ifnar1^KO, Klrk1^KO, Sting^KO, Batf3^KO, Ticam1^KO, Myd88^KO, Ccr5^KO, Cxcr3^KO, CD11c-DTR, and ROSA^DTR C57BL/6 mice were purchased from the Jackson Laboratory. Single-KO mice were bred to generate double-KO mice, including Ticam1^KO Myd88^KO, Ccr5^KO Cxcr3^KO, and Ifnar1^KO Cxcr3^KO. Ncr1^iCre mice (a gift from Eric Vivier, Aix Marseille University Hospital, Marseille, France) were bred with ROSA^DTR mice to generate Ncr1^iCre ROSA^DTR mice. MMTV-PyMt^tg mice (a gift from David DeNardo, Washington University, St. Louis, Missouri, USA) were maintained on BALB/c background. All mice were housed in pathogen-free facilities in accordance with the guidelines instituted by the animal study committees of Massachusetts General Hospital.

C57BL/6, Batf3 KO and Caspase 1 KO mice were purchased from Jackson Laboratories. RIPK3 KO (Ripk3^-/-) (Newton et al., 2004 (link)) and RIPK3 kinase-inactive (Ripk3^K51A/K51A) (Mandal et al., 2014 (link)) were kindly provided by E. Mocarski (Emory University). Tissues from immunized CD169-DTR and LysM-iDTR mice were kindly provided by M. Merad (Icahn School of Medicine at Mount Sinai). ASC KO mice and MLKL KO mice were originally obtained from V. M. Dixit (Genentech) and bred under specific pathogen-free conditions. Mice were immunized subcutaneously at the base of tail with endotoxin-free Ovalbumin (Hyglos) and following adjuvants: MF59 (Novartis), Addavax (Invivogen) and Alum (Alhydrogel; Invivogen). In case of co-administration of cell death inhibitors such as z-VAD-fmk (500 μg/mouse) and Nec-1s (100 μg/mouse) or enzymes including DNaseI (2,000U/mouse), those reagents were mixed with vaccine inoculums right before the subcutaneous injections. In order to deplete LN-resident macrophages, 0.25 mg (50 μl) of Clodrosome (CLL) and Encapsome (control liposome) were subcutaneously injected 5 days prior to each vaccination. Mice were maintained under specific-pathogen-free conditions in the vivarium of Emory Vaccine Center. All animal studies were conducted by following animal protocols reviewed and approved by the Institutional Animal Care and Use Committee of Emory University.

All mice were on a C57BL/6 background. Wild-type (WT) mice (aged 6–8 weeks) were purchased from Daehan Bio Link. Batf KO, Batf3 KO, and Rag1 KO mice were purchased from the Jackson laboratory. All mice were raised under specific pathogen-free conditions, and all animal experiments were approved by the Sogang University Institutional Animal Care and Use committee.