Extended Data Figure 4.
Exogenously added α-KG and 2-HG rescued the effects of AOA on TH17 and iTreg cell differentiation related to Fig. 2f and 2g. a), Cell-permeable dimethyl esters of succinate, fumarate, malate, citrate, NAC or GSH, did not rescue the inhibitory effects of AOA on TH17 cell differentiation. Cell-permeable metabolites (0.75 mM α-KG, 0.5 mM 2-HG, 0.5 mM succinate, 50 μM fumarate, 0.5 mM malate, 0.5 mM citrate, 1 mM NAC, and 1 mM GSH) were individually added to differentiating TH17 cells in the presence of AOA. At the end of differentiation (day 6), the cells were re-stimulated and analyzed by intracellular staining of FOXP3 and IL-17. b) Cell-permeable dimethyl esters of α-KG, 2-HG, but not succinate, fumarate, malate or citrate rescued the effects of AOA on iTreg cell differentiation. Cell permeable metabolites (0.5 mM succinate, 50 μM fumarate, 0.5 mM malate, 0.5 mM citrate) were individually added to differentiating iTreg cells in the presence of AOA. At the end of differentiation (day 5), the cells were directly analyzed for FOXP3-GFP. The representative flow data from 3 independent experiments were shown in a and b (left panel). Bar graphs in the right panel of a and b are mean±S.D. of 3 independent experiments. NS=non-significant; *P<0.05; **P<0.01; ***P<0.001 by Student`s t-test.