Figure 2.
CRBN loss augments energetics of activated CD8+ T cells. (A) Relative expression of Kcna3 (vs B2M) after a 24-hour stimulation of Crbn+/+ and Crbn−/− CD4+ and CD8+ T cells with anti-CD3ε, with or without anti-CD28. (B) Pathway analysis of transcripts that specifically change in activated Crbn−/− T cells (Figure 1D). Basal ECARs (C) and OCRs (D) in anti-CD3ε, with or without anti-CD28–activated Crbn+/+ and Crbn−/− CD8+ T cells. (E-F) Glucose uptake of activated Crbn+/+ and Crbn−/− CD8+ T cells with anti-CD3ε, with or without anti-CD28, as measured by the fluorescent glucose analogue 2-NBDG; G-MFI, geometric MFI; FMO, fluorescence −1. (G) Hexokinase (HK) enzymatic activity of activated Crbn+/+ and Crbn−/− CD8+ T cells. (H) NAD+/NADH ratio of activated Crbn+/+ and Crbn−/− CD8+ T cells. (I) ATP production of activated Crbn+/+ and Crbn−/− CD8+ T cells. (J) Crbn+/+ and Crbn−/− CD8+ T-cell proliferation after 72 hours of anti-CD3ε+anti-CD28 stimulation with 2-deoxyglucose to suppress glycolysis. All results are representative of at least 2 independent experiments. n.s., not significant; * P < .05; ** P < .01; *** P < .001; **** P < .0001.