Figure 2

(A) Staining procedure of STAT5. Whole blood (100 μl) is stimulated by IL-2, IL-7 or IL-15 for 30 min at 37°C. Red blood cells are lysed and white blood cells are fixed for 10 min at 37°C with Lyse/Fix Buffer (BD Biosciences, San Jose, CA). Next, the cells are washed in FacsFlow buffer (BD Biosciences) and permeabilized with cold 70% methanol for 30 min at −20°C, washed twice in FacsFlow buffer supplemented with 0.5% bovine serum albumin followed by standard staining procedures for P-STAT5 (clone Y694), CD3, CD4 and CD8. (B) Whole blood stimulated for 30 min with IL-2, IL-7 and IL-15 (2,000 IU/ml) in the absence and presence of different concentrations of tofacitinib/CP-690,550. P-STAT5 expression by CD3 T cells.