Figure 5: Lkb1 prevents STAT4 binding to CNS2.
(a) Binding of STAT4 and STAT5 to the Foxp3 locus in Foxp3Cre and Foxp3CreLkb1f/f Treg cells treated with IL-2 or IL-2 plus IL-12, determined by ChIP. (b) Binding of Dnmt1 to the CNS2 of Foxp3 locus in Foxp3Cre and Foxp3CreLkb1f/f Treg cells treated with IL-2 or IL-2 plus IL-12, determined by ChIP. (c) STAT4 and STAT5 coprecipitation with Dnmt1 and Dnmt3a was analysed by using nuclear extract from in vitro expanded Treg cells. (d) Methylation of the CpG motifs of the Foxp3 promoter and CNS2 in Foxp3Cre and Foxp3CreLkb1f/f Treg cells determined by bisulfite sequencing, and Treg cells were treated with indicated reagents. Filled circles represent methylated CpG sites and open circles represent unmethylated CpG sites. (e) Foxp3 expression in Foxp3Cre and Foxp3CreLkb1f/f Treg cells co-cultured with DCs supplemented with IL-2 plus IL-12 in the presence or absence of 5-aza-dC. All data are representative of at least two independent experiments.
