Ot 2 rag

C57BL/6 (B6) mice homozygous for loxP flanked caspase-8 allele (Casp8^fl/fl) (21 (link)) were crossed with mice expressing Cre under control of the CD11c promoter (Cre^CD11c, Jackson Laboratory, Alexander Chervonsky), generating Cre^CD11cCasp8^fl/fl mice. PCR on FACS-sorted splenic conventional DC populations (B220⁻CD11c⁺CD8⁻ and B220⁻CD11c⁺CD8⁺) from Cre^CD11cCasp8^fl/fl mice showed deletion of caspase-8 but not in plasmacytoid DCs (CD11c^intermediatePDCA-1⁺B220⁺), lymphocytes or macrophages (Supplemental Figure 1). Cre^CD11cCasp8^fl/fl BMDCs showed caspase-8 deletion (Supplemental Figure 1). OT-II/RAG^−/− and B6.CD45.1 were purchased (Jackson Laboratory). RIPK3^−/− (Genentech), IRF3^−/− (a gift from Mike Diamond), IRF7^−/− (a gift from Mike Diamond), MyD88^fl/fl (Jackson Laboratory) mice were bred to Cre^CD11cCasp8^fl/fl generating RIPK3^−/−Cre^CD11cCasp8^fl/fl, IRF3^−/−Cre^CD11cCasp8^fl/fl, IRF7^−/−Cre^CD11cCasp8^fl/fl and MyD88^fl/flCre^CD11cCasp8^fl/fl mice. Real-time PCR performed by Transnetyx on FACS-sorted splenic conventional DC populations from MyD88^fl/flCre^CD11cCasp8^fl/fl mice showed caspase-8 and MyD88 deletion (Supplemental Figure 1). Female mice were used in all studies. Proteinuria was assessed using uristix (Siemens). Transnetyx performed genotyping. Experiments were approved by Northwestern University IACUC.