Clone mp2 8f8

For cocultures, 100 000 BMMCs were seeded with 25 000 MHC‐II⁻CD4⁺CD25⁺T_reg in the presence of 1.5 µg/ml soluble anti‐mouse CD3ε (clone 2C11, Institute for Immunology) in the presence of IL‐33 (50 ng/ml, Peprotech), with or without IL‐3 (50 ng/ml, Peprotech), with or without SCF (50 ng/ml, Peprotech) in a 96‐well plate for 3 days. In some experiments (as indicated in the figures), we added recombinant IL‐2 (50 ng/ml, Peprotech), a blocking anti‐ICOS‐L antibody (10 µg/ml, clone MIL‐5733²⁷), a blocking anti‐IL‐3 antibody (10 µg/ml, clone MP2‐8F8, Biolegend), or a blocking IL‐6 antibody (10 µg/ml, clone MP5‐20F3, Biolegend) to the cocultures. Coculture experiments were performed with biological replicates (indicated in the figures).

Aged Lyn^–/– mice (min. 25 weeks old) were injected intraperitoneally with 100 µg of isotype control (rat IgG1,κ; clone HRPN, BioXcell), 50 µg^{43 (link)} of rat anti-mouse IL-3 (clone MP2-8F8, Biolegend) or 100 µg^{70 (link)} of rat anti-mouse IL-4 (clone 11B11, Biolegend) 72, 48 and 24 h before analysis.