Anti nk1.1 clone pk136

Isolation of single-cell suspension from the spleens of 8-wk-old B6.CD45.1 mice was performed as previously described (Rose et al., 2012 (link)). Splenic lymphocytes were purified by negative immune-magnetic selection. In brief, spleen cells were treated with Fc Block and incubated with PE-conjugated myeloid-linage antibodies including anti-Ter119 (clone TER119; eBioscience), anti-CD11b (clone M1/70; BD Biosciences), anti-F4/80 (clone BM8; BioLegend), anti-CD11c (clone HL3; BD Biosciences), anti–mPDCA-1 (clone JF05-1C2.4.1; Miltenyi Biotec), anti-SiglecF (clone E50-2440; BD Biosciences), anti-NK1.1 (clone PK136; BD Biosciences), and anti-Ly6G (clone 1A8; BD Biosciences) followed by incubation of anti-PE MicroBeads (Miltenyi Biotec) and depletion on autoMACS Pro Separator (Miltenyi Biotec) with the “depletes” program. The resulting lymphocyte preparation was >80% purity as determined by flow cytometry. Each CD45.2. Rag^−/− or CD45.2. Rag^−/−LysM^CreBim^fl/fl mouse received ∼4.0 × 10⁷ to 5.0 × 10⁷ donor lymphocytes through retro-orbital injections every month over an 8-mo period and bled every month to ensure the efficacy of adoptive transfer. The recipient CD45.2. Rag^−/− and CD45.2. Rag^−/−LysM^CreBim^fl/fl mice were then sacrificed after 8 mo of adoptive transfer to assess SLE-like disease symptoms.