Cd3ε clone 17a2

For samples not sorted, cells were washed with PBS and stained in FACS buffer (2% FBS, 2 mM EDTA, and 0.001% NaN₃). All gating strategies are represented in Supplementary Fig. 9. Cells were first stained for H-2K^b/SIY-pentamer (PE; ProImmune) for 10 min at room temperature at a 1:20 dilution, followed by staining with remaining antibodies for 20 min on ice. Antibodies against the following molecules were used: CD3ε (clone 17A2, BioLegend, 100216), Thy1.2 (clone 30-H12, BioLegend, 105320), CD45.2 (clone 104, BioLegend, 109806), CD8α (clone 53-6.7, BioLegend, 100747), CD4 (clone RM4-5, BioLegend, 100547), PD-L1 (clone 10 F.9G2, BioLegend, 124312), CD19 (clone 6D5, BioLegend, 115545), I-A/I-E (clone M5/114.15.2, BioLegend, 107630), and H-2K^b (clone AF6-88.5, BioLegend, 742862). All antibodies were used at a 1:100 dilution. Fixable Viability Dye eFluor 506 or 780 (eBioscience) was used for live/dead discrimination and was used at a 1:200 dilution. All flow cytometric analysis was conducted on either an LSRFortessa or X20 (BD) and analyzed using FlowJo software (Tree Star).

Anti–DNP IgE mouse mAb (clone ɛ26) (69 (link)) was provided by Fu-Tong Liu (UC Davis Department of Dermatology, Davis, California, USA). mAbs specific for mouse B220 (clone RA3-6B2) Pacific blue–conjugated, CD3ε (clone 17A2) Pacific blue–conjugated, CD11c (N418) FITC–conjugated, CD16/32 (clone 93) purified, CD49b (clone DX5) PerCP-Cy5.5–conjugated, CD117 (clone 2B8) PE–conjugated, and Ly-6G/Ly-6C (clone Gr-1; RB6-8C5) Pacific blue–conjugated purified mouse IgE k isotype control, as well as purified mouse IgG1 k isotype control, were from BioLegend. mAbs specific for mouse IgE (clone R35-72) FITC–conjugated, CD11b (clone M1/70) PE–conjugated, Siglec-F (clone E50-2440) PerCP-Cy5.5–conjugated, IgE (clone R35-72) purified, IgE (clone R35-72) Biotin–conjugated, IgG1 (clone A85-3) purified, and IgG1 (clone A85-1) Biotin–conjugated antibodies, as well as Streptavidin-HRP, were from BD Biosciences. Recombinant Murine IL-3 was from PeproTech. Albumin DNP-HSA; 3, 3′, 5, 5′-Tetramethylbenzidine Liquid Substrate, Supersensitive, for ELISA; Minimum Essential Medium Eagle; PIPES; and Tyrode’s Salt were from Sigma-Aldrich.