Efluor fixable viability dyes

Lung and spleen tissue cells were analyzed using flow cytometry as described previously^{24 (link)}. Briefly, cell samples were incubated in staining buffer with 10% goat serum and 5 µg/mL anti-CD16/CD32 to block nonspecific binding. Antibodies to IL-13, IFN-γ, FoxP3, CD11c, MHC class II, CD4, CD11b, and CD45R/B220 were purchased from eBioscience (San Diego, CA). Antibodies to CD103, CD40, Siglec-F, CD86, and CD45 were from BD Biosciences (San Jose, CA). Anti-neutrophil antibody (7/4) was purchased from Abcam (Cambridge, MA). SHIP-1 antibody (P1C1) was from Santa Cruz Biotechnology (Dallas, TX) and conjugated to PE or Alexa Fluor 647 (AbLab, Vancouver, British Columbia, Canada). ILC2 cells in lung and spleen were identified as Lineage⁻B220⁻CD127⁺CD25⁺CD90.2⁺T1/ST2⁺ cells, which are primarily Sca-1⁺ and CD117⁺. Intracellular staining was performed as described previously with modifications^{24 (link)}. Dead cells were excluded using eFluor fixable viability dyes (eBioscience). Samples were acquired on a BD LSR II Flow Cytometer, and data analysis was performed using the FlowJo software (Tree Star, Ashland, OR).