Anti gata3 bv421

HVS-T cells from six healthy donors and P were cultured in the presence of IL-2 at 20 IU/ml (Roche). HVS-T cells were harvested, counted, and topped up to 10⁶ cells/ml without recombinant IL-2. 200 µl cells per well were plated to 96-well round bottom tissue-culture plates. After 24 h culture, cells were stimulated with 25 ng/ml PMA and 0.5 µM Ionomycin for 3 h. Protein transport inhibitor (eBioscience) was added to each well. After another 3 h, cells were harvested for surface staining with FcBlock, anti-CD271-FITC Abs, and Zombie-NIR live-dead exclusion dye (BioLegend). Cells were then fixed, permeabilized with FOXP3/perm kit (Thermo Fisher Scientific), and subjected to overnight ICS with anti-GATA3-BV421 (BD Biosciences), anti-IL-13-BV711 (BD Biosciences), anti-IFN-γ-eFluor450 (eBioscience), anti-IL-9-PERCP/Cy5.5 (BioLegend), anti-IL-5-PE (BioLegend), anti-T-bet-PE/Cy7 (BioLegend), anti-IL-10-PE/Dazzle594 (BioLegend), anti-IL-22-APC/Fire750 (BioLegend), anti-TNF-BV510 (BioLegend), anti-RORγ/RORγT-BV650 (BD Bioscience), anti-IL-17A-BV785 (BioLegend), and anti-IL-4-Alexa 647 (BioLegend), followed by flow cytometry analysis with an Aurora cytometer (Cytek).