Fig. 8: NAT10 overexpression restores proliferation and antiviral defects in T cells from older animals.
a, NAT10 and MYC overexpression in T cells from aged mice. Representative data from three independent experiments are shown. Y, young; O, old; w, weeks. b,c, CellTrace dilution analysis (b) and Ki-67 expression (c) in T cells from aged mice with NAT10 or MYC overexpression. Left, representative flow plots. Right, bar diagrams showing the percentage of CellTracelo (b) or Ki-67+ (c) T cells in each group; n = 5 (b) and n = 4, 3, 4 and 4 (c). Exact P values from left to right: **P = 0.0052, *P = 0.0315 and **P = 0.0065 (b) and ****P < 0.0001, ***P = 0.0005 and ***P = 0.0001 (c). d–f, Glucose consumption (d), lactate production (e) and ATP generation (f) in T cells from aged mice with NAT10 or MYC overexpression; n = 5, 6 and 4. Exact P values from left to right: *P = 0.0115, **P = 0.0048 and *P = 0.0122 (d), ***P = 0.0001, ***P = 0.0001 and ***P = 0.0006 (e) and **P = 0.0081, **P = 0.0086 and ****P < 0.0001 (f). g, OPP staining of T cells from aged mice with NAT10 or MYC overexpression. Left, representative flow plots. Right, bar diagrams showing the percentage of OPP+ T cells in each group; n = 3; ***P = 0.0010, **P = 0.0073 and *P = 0.0428. h, Flow chart of experiments exploring the impact of NAT10 restoration on T cell antiviral potency. CD8+ T cells from old P14 mice with control or Nat10-overexpressing plasmids were transferred into 8-week-old Rag2−/− mice 1 day before LCMV infection. Eight days later, antiviral analysis was performed. i, Serum viral loads of recipients at 8 days after LCMV infection; NC n = 6, Nat10 overexpression (OE) n = 5; *P = 0.0424. j, Proportions of GFP+ P14 T cells in recipients. Left, representative flow plots. Right, bar diagrams showing the expansion fold changes of GFP+ P14 T cells in each group; NC n = 6, Nat10 overexpression n = 5; *P = 0.0195. k, Detection of IFNγ+TNF+GFP+ CD8+ T cells in recipients. Left, representative flow plots. Right, bar diagrams showing the absolute number of splenic IFNγ+TNF+GFP+ CD8+ T cells in the two groups; NC n = 6, Nat10 overexpression n = 5; *P = 0.0415. n refers to the number of biologically independent samples. Error bars represent mean ± s.e.m. (b–g and i–k). Data were analyzed by two-tailed, one-way ANOVA with a Tukey’s multiple comparisons test (b–g) or two-tailed, unpaired t-test (i–k).
