Figure 7.
PTEN deletion leads to Treg instability in vitro. (a) CD4+Foxp3+CD25+ iTregs from Ptenfl/fl Foxp3GFP-hCre mice with indicated genotypes were sorted and cultured in the presence of IL-2 with or without the PI3K inhibitor (10 μM LY294002) for 6 d and expression of Foxp3 and CD25 was monitored at indicated times. Representative of 3 experiments. (b) Foxp3+ cells were isolated from wild-type and Pten-ΔTreg mice, serum starved for 1 h and stimulated for 30 min in the presence of 200 IU/ml hIL-2. Cells were then immediately fixed, permeabilized and assessed for pSTAT5 levels by phospho-flow cytometry. Grey = isotype control. Representative of 3 experiments.