Figure 7. mTORC1-AMPK signaling regulates both glucose and glutamine metabolism in NKT cells.

(A–D) Sorted splenic NKT cells were stimulated for 3 days in the presence or absence of rapamycin (2 nM). Representative histograms and summary graphs show relative HK2 expression (A), CD98 expression (B), GSH production (C), and O-GlcNAc levels (D) in stimulated NKT cells (n = 3).

(E and F) NKT cells were activated for 3 days under the indicated conditions, and relative expression levels of pS6^Ser235/236 (E) and c-Myc (F) were compared (n = 3).

(G) Relative expression of pAMPK before (day [D] 0) and after activation for 3 days (D3) is shown (n = 3).

(H) Cell survival and proliferation of WT and AMPK KO splenic NKT cells after 3 days of activation are shown (n = 3).

(I–K) WT and AMPK KO NKT cells were sorted and stimulated for 3 days. Glutamate and αKG (I), GlcNAc (J), and L-PHA (K) expression levels were compared (n = 3).

(L) Graphs show cytokine expression in WT and AMPK KO NKT cells after 3 days of activation (n = 3).

(M) Histograms show cell proliferation in WT and AMPK KO NKT cells activated with and without CB839 (250 nM) (n = 3). All relative levels were calculated using the average control value as a reference point. All data are combined from at least three independent experiments.

Data are shown as mean ± SEM. *p < 0.05, **p < 0.01.