Anti mouse foxp3 clone mf23

T cells and their subsets were analyzed by differential expression of surface markers as well as by their intracellular cytokine profile. Briefly, cells were stained with different surface markers, such as anti-mouse CD3e (clone BM10-37, BD Biosciences), anti-mouse CD4 (clone RM4-5, BD Biosciences), anti-mouse CD8a (clone53-6.7, BD Biosciences) and anti-mouse CD25 (clone PC61, BD Biosciences). For analysis of the expression of intra cellular cytokines, cells were stimulated with 50 ng of PMA and 1 μg of ionomycin for 4 h in the presence of Golgi stop (BD Biosciences) to inhibit the secretion of the cytokines from the cells. Cells were then washed with staining buffer, fixed and permealized with BD cytofix and cytoperm^TMPlus (BD Biosciences) along with 0.2% Tween 20 for 20 minutes. Next, cells were stained with anti-mouse IL4 (clone 11B11, BD Biosciences), anti-mouse IFN-γ (clone XMG1.2, BD Biosciences), anti-mouse IL-17A (clone TC11-18H10) and anti-mouse Foxp3 (clone MF23, BD Biosciences). Brilliant stain buffer (BD Biosciences) and ultra comp beads (eBioscience, San Diego) were used for the dilution of the stains and compensation respectively. All the stained cells were analyzed by LSRII flow cytometer (BD Biosciences) and the data was evaluated by FlowJo software.