Pe conjugated anti il 13 ebio13a

For co-culture experiments, MLN cells from IL-33-treated mice were stained with PerCp-Cy5.5-conjugated CD90.2 (30-H12) and APC-Cy7-conjugated CD45 (30-F11, Biolegend, San Diego, CA) followed by cell sorting with a FACSAria II (BD Biosciences, San Jose, CA) For intracellular cytokine analyses, after restimulation with phorbol 12-myristate 13-acetate/ionomycin and treatment with Golgi blocker, LP and MLN cells were stained with biotinylated anti–T1/ST-2 (DJ8, MD Biosciences, Oakdale, MN) followed by Streptavidin-BV650, APC-conjugated anti-CD4 (GK1.5), FITC-conjugated anti-CD3 (17A2) and PerCp-Cy5.5-conjugated B220 (RA3–6B2), FCεRI (MAR-1), CD11c (N418), NK1.1 (PK136) and CD11b (M1/70) (Biolegend). Stained cells were fixed and permeabilized for intracellular cytokine staining by using PE-conjugated anti–IL-13 (eBio13A, eBioscience, Waltham, MA). MLN and LP cells were then analyzed with an LSR II (BD Biosciences, San Jose, CA).