Isolation of Hepatic ILC2s from Mice

To expand hepatic ILC2s for isolation, C57BL/6 mice were treated with rmIL-33 (0.3 µg/mouse) on four consecutive days. Hepatic non-parenchymal cells were isolated by Percoll density gradient centrifugation. Single non-parenchymal cells were stained with lineage antibodies (anti-CD3, 145-2C11; anti-CD11b, M1/70; anti-CD45R/B220, RA3-6B2; anti-TER-119, TER-119; anti-GR1, RB6-8C5; all APC; all BioLegend), anti-Sca-1 (D7, Pacific Blue) and anti-ST2 (RMST2-2, PE-Cy7; both BioLegend) antibodies. Lineage-negative (lin^-) cells were enriched by MACS and ST2⁺ Sca-1⁺ lin^- ILC2s were isolated by FACS (BD FACSAriaTM III sorter; BD Biosciences, Heidelberg, Germany) (23 (link)).