Anti ter119 clone ter119

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.

For FACS sorting and analysis, we used previously described staining protocols (Mossadegh-Keller et al., 2013 (link)), published stem and progenitor cell definitions (Bryder et al., 2006 (link)), FACSCanto, LSRII, and FACSAriaIII equipment, and DIVA software (BD), analyzing only populations with at least 200 events. The following antibodies were used for staining cells: anti-CD117 (clone 2B8; BD), anti–Sca-1 (clone D7; BioLegend), anti-CD34 (clone RAM34; BD), anti-CD16/32 (clone 2.4G2; BD), anti-CD11b (clone M1/70; BD), anti-CD19 (clone 1D3; BD), anti-CD3e (clone 145-2C11; BioLegend), anti-Ly6G (clone 1A8; BioLegend), anti-Ly6C (clone HK1.4; BioLegend), anti-CD115 (clone AFS98; eBioscience), anti-CD45.2 (clone 104; BD), anti-CD45.1 (clone A20; BD), anti-B220 (clone RA3-6B2; eBioscience), anti-Ter119 (clone TER-119; eBioscience), and anti-CD71 (clone R17217; eBioscience). LIVE/DEAD Fixable Violet Dead cell dye (Invitrogen) was used as a viability marker. Information describing gating strategies is shown in Figs. S1 and S2.