Cells were immunostained with the following monoclonal anti-human antibodies: FITC anti-Lin3, V450 anti-CD56, APC-H7 anti-CD45, APC anti-IFN-γ, PE anti-TNF-α, PE Annexin V, 7AAD (all from Becton Dickinson, Milan, Italy), PercpCy5.5-CD127, PE anti-TLR3, PE anti-TLR9, APC anti-TLR4, PE anti-TLR2, APC anti-ROR-γt, PE anti-ROR-γt, PECy7 anti-T-bet, PE anti-IL-17A, AlexaFluor647 anti-IL-17A (all from eBioscience), PE anti-CRTH2 (Biolegend, San Diego, CA) and PE anti-TLR7 (R&D Systems). Cells were immunostained with the following anti-mouse antibodies: FITC antiCD90.2, hematopoietic lineage cocktail efluor 450, PECy7 anti-T-bet, APC anti-ROR-γt, PE anti-TNF-αPE anti-IL-17A (all from eBioscience), APC-Cy7 anti-CD45, PE anti-ROR-γt, PECy7 anti-IFN-γ (all from Becton Dickinson). In all experiments, appropriate isotype control IgGs (Becton Dickinson and eBioscience) and fluorescence minus one controls were used. All antibodies were used at 1:100 final dilution. For intracellular immunostaining, cells were fixed and permeabilized using staining buffer set and permeabilization buffer (both from eBioscience) according to the manufacturer’s instruction. Cells were analyzed by flow cytometry (FACSverse, BD Bioscience, San Jose, CA).
Marafini I., Monteleone I., Di Fusco D., Cupi M.L., Paoluzi O.A., Colantoni A., Ortenzi A., Izzo R., Vita S., De Luca E., Sica G., Pallone F, & Monteleone G. (2015). TNF-α Producing Innate Lymphoid Cells (ILCs) Are Increased in Active Celiac Disease and Contribute to Promote Intestinal Atrophy in Mice. PLoS ONE, 10(5), e0126291.
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