Flow Cytometry
Single cell suspensions were incubated with Aqua Live/Dead Fixable Dye
(Life Technologies, Grand Island, NY) an fluorochrome-conjugated
monoclonal antibodies (mAbs) against mouse CD3 (17A2), CD64 (X54-5/7.1),
CD5 (53-7.3), CD11c (N418), CD19 (eBio1D3), NK1.1 (PK136), TCRγδ
(eBioGL3), CD45 (30-F11), IL-33R/ST2 (RMST2-2), CD49b (DX5), FcεRIα
(MAR-1), IgE (R35-72), and Notch2 (16F11) from Thermo Fisher Scientific,
BD Biosciences, or BioLegend. Staining was performed in the presence of
10% rat serum (Jackson ImmunoResearch) and 1 μg/mL FcR block
(anti-CD16/32, 2.4G2; Bio X Cell). Stained cells were acquired on an
LSRII (BD biosciences) and analyzed using FlowJo software version 10.8.