Anti cd90.2 pe cy7

Cells were incubated with an anti-CD16/CD32 monoclonal antibody (mAb, eBioscience, 14-0161-82, 93, 1:33) and then stained with the following dye or anti-mouse antibodies: viability dye APC-Cy7 (eBioscience, 65-0865-14), anti-lineage cocktail-FITC (BioLegend, 133302, 145-2C11; RB6-8C5; RA3-6B2; Ter-119; M1/70, 1:40), anti-TCRβ-FITC (BioLegend, 109206, H57-597, 1:200), anti-CD90.2-PE/Cy7 (BioLegend, 105326, 30-H12, 1:200), anti-CD127-APC (BioLegend, 135012, A7R34, 1:100), anti-ST2-BV421 (BioLegend, 145309, DIH9, 1:100), anti-CD3E-FITC (BioLegend, 100203, 17A2, 1:200), anti-TCRg-PE (BioLegend, 118107, GL3, 1:100). Gating was performed as follows: ILC2s were defined as live Lin⁻ (CD3e⁻Ly6G⁻Ly6c⁻CD11b⁻CD45R⁻TER119⁻TCRb⁻) CD127⁺Thy1⁺ST2⁺ cells. Stained cells were examined with a FACSCanto II flow cytometer (BD Biosciences), and the results were analyzed using FlowJo software version 10.4.2 (TreeStar, FlowJo).

Immune cells from the colon were isolated according to recent protocol^{63 (link)}. The isolated immune cells were resuspended in FACS buffer (2% FCS in PBS) and were passed through a 70-μm strainer (BD Biosciences). Samples were analysed with following antibodies: anti-CD11b-PE (1:100 dilution, 101208, Biolegend), anti-CD11c-PE-Cy7 (1:100 dilution, 117318, Biolegend), anti-γδTCR-APC (1:100 dilution, 118116, Biolegend), anti-γδTCR-PE-Cy7 (1:100 dilution, 118123, Biolegend), anti-βTCR-FITC (1:100 dilution, 109205, Biolegend), anti-βTCR-PE (1:100 dilution, 553172, BD Biosciences), anti-CD45-V510 (1:100 dilution, 103137, Biolegend), anti-CD45-PE-CF594 (1:200 dilution, 562420, BD Biosciences), anti-CD45-APC (1:200 dilution, 103112, Biolegend), anti-CD90.2-PE-Cy7 (1:200 dilution, 140309, Biolegend), anti-CD3-PE-CF594 (1:200 dilution, 562332, BD Biosciences), anti-F4/80-APC-Cy7 (1:50 dilution, 123119, Biolegend), anti-CD19-FITC (1:100 dilution, 115505, Biolegend), anti-CCR-6-V421 (1:50 dilution, 129817, Biolegend), anti-IL-6R-PE (1:50 dilution, 115805, Biolegend) and LIVE/DEAD Fixable Violet Dead Cell Stain Kit or LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (1:1000 dilution, L34955 and L34957, ThermoFisherScientific). Data were acquired on MACSQuant Analyzer (Miltenyi) and MACSQuant VYB (Miltenyi) and data were analysed with FlowJo software (Treestar).