Il 13 clone ebio13a

Peptide restimulation and ICS for CD8⁺ T cells were performed as previously described (1 (link)). For CD4⁺ T cell and innate lymphoid cell ICS, lung cells were stimulated with PMA (50ng/ml) and ionomycin (1ug/ml) (Sigma) in the presence of 0.067% GolgiStop (BD) for 5 hours. After stimulation, cells were stained for viability (live/dead violet, Life Technologies) CD4 (clone RM4-5, eBioscience), IFNγ (clone XMG1.2, BD), and IL-4 (clone 11B11, Biolegend), or ILC markers (for Fig 6F, pan-ILC markers CD45, CD127, Lineage, CD90; for all ILC2 ICS figures, pan-ILC markers plus ICOS)(see above for clones) and IL-5 (clone TRFK5, BD), IL-13 (clone eBio13A, eBioscience) and IFNγ (clone XMG1.2, BD).

Cell suspensions were stained in ice cold PBS with or without 2 % FBS on ice. The following antibodies were used for staining mouse cells: CD45 (clone 30-F11, Biolegend), Ly6G (clone 1A8, BD Biosciences), CD11c (clone HL3, BD Biosciences), CD11b (clone M1/70, Biolegend), Siglec F (clone E50–2440, BD Biosciences), CD3e (clone 145–2C11, eBioscience), CD4 (clone GK1.5, Biolegend), IFN © (clone XMG1.2, Biolegend), IL-17 (clone eBio17B7, eBioscience), IL-5 (clone TRFK5, Biolegend), IL-13 (clone eBio13A, eBioscience), CD25 (clone PC61.5, eBioscience), CD44, CD62L (clone MEL-14, BD Biosciences), MHC class II I-A/I-E (clone M5/114.15.2, eBioscience), DC-SIGN/CD209 (R&D systems), CD32 (clone D34–485, eBioscience), TLR4/CD284 (clone SA15–21, Biolegend), Annexin V (Biolegend). Neutrophils were defined as CD45⁺CD11b⁺Ly6G⁺DNA⁺ and cytoplasts were defined at CD45⁺CD11b⁺Ly6G⁺DNA⁻. Naïve T lymphocytes were defined as CD4⁺CD25⁻CD44^lowCD62L^hi. Lung Dendritic cells were defined as CD45⁺CD11c⁺ MHCII⁺autofluorescence^low. The following antibodies were used to stain human cells: anti-CD45 PE-Cy7 (HI30), anti-CD66b (G10F5), anti-CD16 APC-Cy7 (3G8) all from Biolegend. Vybrant DyeCycle Ruby (Life Technologies) was used to stain intracellular DNA. Data were acquired on BD Canto II or BD LSR Fortessa and analyzed using FlowJo v10. For cell sorting, FACS Aria was used.