Extended Data Fig. 5: Met limitation results in increased TCR-mediated NFAT1 activation.

a, Unedited image of Fig. 3d showing the unmasked, red anti-CD3/38 Dynabeads which are masked grey in the main Figure to highlight NFAT1 staining. b, NFAT1 intensity quantification (nuclear to total cell ratio) of CD8⁺ T cells activated in either 0.1 mM, 0.00 mM or 0.03 mM Met for 30 min with anti CD3/28 Dynabeads and stained for NFAT1 (each circle represents one cell, n = 28 cells/group). c, Quantification of nuclear NFAT1 from the experiment shown in Fig. 3d. d, NFAT1 intensity quantification (nuclear to total cell ratio) of CD8⁺ T cells activated for 30 min by CD3/28 Dynabeads in either 0.1 mM or 0.03 mM Met in the presence of DMSO or 5 μM YM58483 (each circle represents one cell, n = 47 cells/group). e, NFAT2 quantification (nuclear to total cell ratio) of CD8⁺ T cells, activated in 0.1 mM or 0.03 mM Met for 30 min with anti-CD3/28 Dynabeads (each circle represents one cell, n = 40 cells). f, NFAT1 quantification (nuclear to total cell ratio) of previously activated OT-I T cells, treated with anti-CD3/28 Dynabeads for 30 min. T cells were initially activated with 2.5 ng/ml SIINFEKL for 24 h in control medium and rested with 10 ng/ml mIL-7 for 48 h (each circle represents one cell, n = 50 cells/group). g, Normalized RNA expression quantified by quantitative-PCR performed 24 h post activation of OT-I CD8⁺ T cells activated with 2.5 ng/ml SIINFEKL in 0.1 mM or 0.03 mM Met for 30 min followed by 0.1 mM Met (n = 5–6 biological replicates/group). Data are mean±s.d. Unpaired two tailed Student’s t-test (b–f), paired two-tailed Student’s t-test (g).

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