Fitc anti mouse nk1

DCs were sorted after isolation from spleen or colon from propyzamide- or vehicle-treated mice as described above. Cell suspensions were stained with FITC anti-mouse-NK1.1 (108706, BioLegend), CD64 (139316, BioLegend), Ly6G (127606, BioLegend), Ly6C (128006, BioLegend), B220 (553087, BD Bioscience) F/480 (11-4801-85, Thermo Fisher Scientific), PECy7 or PE-Dazzle594 anti-mouse-CD11c (N418, BioLegend) or PECy7-anti mouse CD3 (100220 and 117348, BioLegend) antibodies. After 30 min on ice, cells were washed and sorted on the FACS Aria Ilu (BD Biosciences) system. T cells were used to reconstituted Rag1-deficient mice as described below. DCs were pulsed with 10 μg ml⁻¹ of OVA 323-339 (SP-51023-1, Genemed) for 6 h, washed and co-cultured with naive CD4⁺ T cells isolated from spleens from B6.Cg-Tg(TcraTcrb)425Cbn/J OT-II mice using magnetic beads (130-104-453, Miltenyi). After 48 h, cells were restimulated and stained as described below.

Extract from Dermatophagoides farinae (Df) was obtained from Greer Laboratories (XPB81D3A25; Lenoir, NC). Ovalbumin (OVA) and PBS were obtained from Sigma-Aldrich (St Louis, Mo). The mMCP-1 EIA kit was purchased from eBiosciences (San Diego, Calif). LTA₄, LTC₄, LTD₄, LTE₄, MK571, and HAMI3379 were obtained from Cayman Chemical (Ann Arbor, Mich). Histamine, thromboxane receptor B₂, PGD₂, and cysLT EIA kits were obtained from Cayman. IL-4, IL-5, IL-13, ICAM-1, and VCAM-1 EIA kits were from R&D Systems (Minneapolis, Minn). The CXCL7 EIA kit was purchased from Abcam (Cambridge, Mass). The HMGB1 EIA kit was from LifeSpan (Providence, RI). The monoclonal goat anti-mouse IL-33 was purchased from R&D Systems (Minneapolis, Minn), and the rat anti-mouse IgG (H1L) secondary antibody, fluorescein isothiocyanate (FITC) anti-mouse CD11c, FITC anti-mouse/human CD11b, FITC anti-mouse IgE, FITC anti-mouse CD3ε, FITC anti-mouse CD19, FITC anti-mouse CD8a, FITC anti-mouse NK-1.1, FITC anti-mouse Ly-6G/Ly-6C (Gr-1), allophycocyanin (APC) anti-mouse CD45, APC/cyanine 7 (Cy7) anti-mouse/human CD44, PerCP/Cy5.5 anti-mouse CD90.2, phycoerythrin (PE) anti-mouse CD278 (inducible costimulatory molecule), APC anti-mouse CD41, PE/Cy7 anti-mouse CD62P, APC anti-human CD61, anti-mouse CD16/32, PE/Cy7 anti-mouse CD45, and PE anti-mouse Siglec F were all obtained from BioLegend (San Diego, Calif).