Extended Data Fig. 4: NAT10 deletion causes metabolic dysfunction of T cells.
a Heatmap summarizing expressions of critical genes in glutaminolysis, glycolysis, amino acids biosynthesis and nucleotide metabolism in CKO versus FLOX TEFF cells from RNA-seq data (related to Fig. 4a). Expression data was Z-score normalized. b Differential activity of indicated metabolic reactions between FLOX and CKO splenic T cells calculated via “Compass” algorithm based on scRNA-seq data. Effect size was assessed with Cohen’s d statistic, namely absolute difference in means divided by the pooled SD. c, d TN were purified from 8-week-old FLOX and CKO mice, part of which were stimulated with anti-CD3/28 for 24 h. Then, FLOX TN, CKO TN as well as their activated counterparts were subjected to metabolic profiling. Heatmap of nucleotide profiles (c), metabolite profiles (d) in FLOX TN, FLOX TEFF, CKO TN and CKO TEFF were presented. Data was Z-score normalized. e TN were purified from 8-week-old FLOX and CKO mice and stimulated with anti-CD3/28 for 24 h. Glucose uptake measured by 2-NBDG fluorescence intensity through FCM was displayed. FLOX n = 5, CKO n = 6. ****P < 0.0001. f Relative ECAR and OCR of FLOX and CKO TN measured at baseline or with anti-CD3/CD28 beads stimulation. For ECAR, FLOX-TN n = 10, CKO-TN n = 9, FLOX-CD3/28 n = 7, CKO-CD3/28 n = 8; ***P (interaction) = 0.0006, P (*marked timepoint) = 0.0092, 0.0081, 0.0022, 0.0043, 0.0051, 0.0096, 0.0020, 0.0043, respectively. For OCR, FLOX-TN n = 9, CKO-TN n = 3, FLOX-CD3/28 n = 5, CKO-CD3/28 n = 4; **P (interaction) = 0.0090, P (*marked timepoint) = 0.0422, 0.0135, 0.0070, 0.0056, 0.0270, 0.0193, 0.0098, 0.0017, respectively. g TN were purified from 8-week-old FLOX and CKO mice, part of which were stimulated with anti-CD3/28 for 24 h. Then, ATP production of FLOX-TN (n = 6), CKO-TN (n = 5) as well as their activated counterparts (n = 5) were measured. **P = 0.0016, ****P < 0.0001. The values of “n” all refer to biologically independent samples. Error bars represent mean ± s.e.m. (e–g). Two tailed, negative binomial distribution test (e); two-tailed, two-way ANOVA with Šídák’s multiple comparisons test (f); two-tailed, one-way ANOVA with Tukey’s multiple comparisons test (g).
