Anti tbet apc

The murine spleen cells and lymph nodes cells were stimulated using PMA, ionomycin and protein transport inhibitor (BD GolgiStop) for 4 h. At the end of stimulation, the cells were permeabilized using IC fixation buffer and 1× permeabilization buffer (eBioscience, San Diego, CA, USA). The following antibodies were used for flow cytometry: anti‐IFN‐γ‐APC (177311, eBioscience, San Diego, CA, USA), anti‐IL‐17A‐PE (130103015, Miltenyi, Auburn, CA, USA), anti‐Tbet‐APC (644813, Biolegend, San Diego, CA, USA), anti‐Foxp3‐FITC (eBio7979; 115773, eBioscience, San Diego, CA, USA), anti‐RORγt‐PE (562607, BD Pharmingen, San Diego, CA, USA), anti‐CTLA4‐PE (121522, eBioscience, San Diego, CA, USA) and anti‐CD4‐VioBlue (130102456, Miltenyi, Auburn, CA, USA). Single‐cell suspensions were examined on a FACS Fortessa (BD Immunocytometry Systems, San Jose, CA, USA), and the data were analysed using FlowJo software. The numbers in the corners of the FACS dot plots represent the percentage of each cell population within that quadrant as a fraction of the total cell population.