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. 2016 Nov 3;11(11):e0165639. doi: 10.1371/journal.pone.0165639

Fig 4. Glycolytic metabolism assessed in competent or NOS2 deficient γδ T cells.

Fig 4

Sorted γδ T cells from pLNs of WT or Nos2KO mice were expanded in vitro for 4 days in presence of CD3- and CD28- specific antibodies, 15 μg/mL IL-7 and 15U/mL IL-2. Glycolytic metabolism was analyzed using a Seahorse XF-24 analyzer either directly (A), or after 18 h of resting followed by an additional 4h of stimulation with media containing 5 mM L-NMMA when indicated (B). ECAR was assessed after glucose (gluc) addition and in response to metabolic inhibitors oligomycin (oligo) and 2-Deoxy-D-glucose (2DG). Shown are time courses (A left), normalized time courses as % of baseline (B left) and calculations of rate of glycolysis (A, B right panels). Data are from one experiment with 3 (Nos2KO) and 4 (WT) replicates (A) and are pooled from three independent experiments with 7 (WT), 5 (Nos2KO) and 5 (L-NMMA) replicates (B). Mean ± SEM are shown. ** p < 0.01 (Mann-Whitney’s test).