Fig. 6
AMPK suppresses the activity of GSK3β through p38 MAPK activation to regulate the expression of PD-1. A Analysis of phosphorylated and total GSK3β and β-catenin in WT and AMPK-KO Tregs stimulated with or without anti-CD3/CD28 stimulation for 4 h. Analysis of the expression of GSK3β and β-catenin by western blotting in activated Tregs after B AICAR and C compound C treatment at the indicated doses. D Immunoblot analysis of the indicated proteins in activated Tregs treated with the p38 MAPK inhibitor SB203580 at the indicated doses. E Immunoprecipitation assay of the lysates of WT Tregs using anti-p38 and anti-GSK3β antibodies. Flow cytometric analysis of F PD-1 and G T-bet in Tregs from WT and AMPKfl/flFoxp3-Cre mice after i.p. treatment with or without the GSK3β inhibitor SB216763 daily for 7 days (2 μg/kg). The data are presented as the mean ± standard deviation (SD); n = 5 mice per group. *P < 0.05; **P < 0.01; ***P < 0.001