Figure 6: Lkb1 suppresses NF-κB signalling to restrain STAT4 activation.

(a) Intracellular phosphorylated NF-κB p65 in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells with or without IL-2 stimulation (n=3). (b) Stat4 and Il12rb2 mRNA in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells supplemented with or without 4-ASA for 16 h (n=3). (c) Intracellular phosphorylated STAT4 in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells supplemented with or without 4-ASA (n=3). (d) P65 enrichment to consensus sequences (CS) in Stat4 and Il12rb2 locus in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells determined by ChIP. (e) Foxp3 expression in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells co-cultured with DCs supplemented with IL-2 and IL-12, with or without 4-ASA. (f) Intracellular phosphorylated IKKα (Ser176/180)/β (Ser177/181) in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells with or without IL-2 stimulation (n=3). (g) Intracellular phosphorylated IκBα (Ser 32/36) in Foxp3^Cre and Foxp3^CreLkb1^f/f T_reg cells with or without IL-2 stimulation (n=3). Two-way analysis of variance (ANOVA) was used for statistical analyses in (a,b,c,d,f,g) (*P<0.05, **P<0.01); error bars represent s.d.; all data are representative of at least two independent experiments.