Pe conjugated anti il 13

The reagents used in this study are as follows: APC- or Bv510-conjugated anti-CD4, FITC- and PE/Cy7-conjugated KJ1-26 (anti-clonotypic for D11.10 TCR), FITC-conjugated anti-CD44, FITC-conjugated anti Lineage Cocktail (containing CD3, Gr-1, CD11b, B220, Ter119), APC- and Bv510-conjugated anti-CD127 (IL-7Rα), APC-conjugated anti-IL-5, PE/Cy7-conjugated anti-Thy1.2, PE-conjugated anti-TCR-beta, PE-conjugated anti-CD335 (Nkp46), Bv421-conjugated ST2, Bv421-conjugated anti-human NGFR and Zombie NIR^TM fixable viability kit were purchased from BioLegend, San Diego, CA.
PE-conjugated anti-CCR6 was purchased from R&D systems. Alexa488-conjugated anti-phospho-p38 was purchased from Cell signaling. PE-conjugated anti-IL-13 was purchased from eBioscience. Anti- PE mouse anti-Human NGFR was purchased from BD Biosciences. Anti-IFNγ monoclonal antibody was purchased from BioLegend. Recombinant mouse IL-4 was from TOYOBO, Osaka, Japan. Recombinant mouse IL-33 and mouse IL-25 were purchased from R&D. Recombinant murine IL-7 was purchased from PeproTech. The OVA peptide (residues #323-339; ISQAVHAAHAEINEAGR) was synthesized by BEX Corporation, Tokyo, Japan. SB203580 (Merck) were used for inhibition of p38 activity. Anti-mouse Thy1.2 antibody was purchased from BioX Cell. Detailed information of antibody is listed in Supplementary Table 4.

Astragalin was modified into astragalin-galactoside (Ast-Gal) using β-galactosidase from Bacillus circulans, and purified by the medium pressure chromatography with silica C-18 column followed by Sephadex LH-20 column [25 (link)]. Ast-Gal was identified by nuclear magnetic resonance to be kaempferol-3-O-β-d-glucopyranosyl-(1->6)-β-d-galactopyranosyl-(1->4)-β-d-galactopyranoside. The water solubility of Ast and Ast-Gal were 28.2 ± 1.2 mg/L and 38,800 ± 2.8 mg/L, respectively. Anti-CD3ε, anti-CD28, and anti-IL-12p70 antibodies were purchased from BD Biosciences (San Diego, CA, USA). Mouse recombinant IFN-γ and IL-4 were purchased from PROSPEC (East Brunswick, NJ, USA). Mouse recombinant IL-6 and TGF-β were purchased from PEPROTECH (Rocky Hill, NJ, USA). FTIC-conjugated anti-CD4, FTIC-conjugated anti-MHC Class II, FTIC-conjugated anti-IgG 2b/k, PE-conjugated anti-IFN-γ, PE-conjugated anti-IL-4, PE-conjugated anti-CD80, PE-conjugated anti-CD86, PerCP-Cy5.5-conjugated anti-CD4, and APC-conjugated anti-IFN-γ antibodies were purchased from BD Biosciences. PE-conjugated anti-Rat IgG2a, PE-conjugated anti-IL-13, APC-conjugated anti-IL-17A, and APC-conjugated anti-CD11c were purchased from eBioscience (San Diego, CA, USA).