Armenian hamster igg isotype ctrl

At indicated times, the mice were fed a HFD (Rodent diet with 60 kcal% Fat, D12492i) from Research Diets Inc. (New Brunswick, New Jersey) for the indicated times^{28 (link)}. Briefly, all other mice were fed a normal chow diet (NCD). For in vivo experiments investigating the effect of DR3 engagement, DR3 agonist (1 mg/mouse, BioLegend, San Diego, CA, clone 4C12), or the monoclonal antibody Armenian Hamster IgG Isotype Ctrl (BioLegend) (1 mg/mouse) was administered intraperitoneally every 4 days from the indicated start of treatment until termination of the experiment.

Murine VAT and human peripheral ILC2s were isolated to >95% purity using the FACS Aria III cell sorter. For in vivo stimulation of murine VAT ILC2s, carrier free rm-IL-33 (Biolegend, San Diego, CA, 1 µg/mouse in 200 µL) was administered intraperitoneally to mice on three consecutive days. On day 4, murine ILC2s were isolated based on the lack of expression of classical lineage markers (CD3ε to mice on three consecutive days. On day TCRγδ and FCεRI) and positive expression of CD45, ST2, and CD117. Isolated ILC2s were stimulated (5 × 10⁴/mL) with rm-IL-2 (10 ng/mL) and rm-IL-7 (10 ng/mL) for 48 h at 37 °C in presence of DR3 agonist (5 µg/mL) from BioLegend, San Diego, CA (clone 4C12, or the monoclonal antibody Armenian Hamster IgG Isotype Ctrl (BioLegend). For adoptive transfer experiments, 2.5 × 10⁵ purified ILC2s were adoptively transferred intravenously in 200 µL PBS into the recipients at the start of the indicated treatment.