Figure 2: GT5 T cells are highly glycolytic in vitro.

A) Schematic for the experiments below.

B) NMR spectroscopy analysis of extracellular [2-¹³C] lactate, produced by OT-I cells following a 4h incubation with [2-¹³C] glucose or [2-¹³C] fructose alone. Peak areas were used for measuring concentrations. Mean lactate concentrations ± SEM shown (n=3, two-tailed Student’s T test).

C) NMR spectroscopy analysis of extracellular ¹³C lactate, produced by OT-I cells, following a 4h incubation with [2-¹³C] glucose plus unlabeled fructose or with [2-¹³C] fructose plus unlabeled glucose. Analysis as in B). (n=3, two-tailed Student’s T test).

D) Heatmap of LC-MS derived fractional enrichment of indicated intracellular metabolites in OT-I cells following a 4h incubation with [U-¹³C] glucose plus unlabeled fructose or with [U-¹³C] fructose plus unlabeled glucose; bold text indicates isotopic. Gradient of blue shading represents the magnitude of fractional enrichment.

E) Fractional enrichment of representative metabolites as in D). n=3, one-way ANOVA.

F) Same as in B) but in 19BBz CAR T cells. n=3, two-tailed Student’s T test.

G) Same as in C) but in 19BBz CAR T cells. n=3, two-tailed Student’s T test.

*, p < 0.05; **, p < 0.01; ***, p < 0.001.

See also Figure S3.