Fig. 3: Loss of NAT10 causes impaired T cell proliferation and enhanced apoptosis. | Nature Immunology

Fig. 3: Loss of NAT10 causes impaired T cell proliferation and enhanced apoptosis.

From: A critical role of N4-acetylation of cytidine in mRNA by NAT10 in T cell expansion and antiviral immunity

Fig. 3

a, Schematic showing the in vivo competitive proliferation assays. TN cells isolated from 8-week-old CD45.1+ FLOX and CD45.2+ CKO mice were labeled with CellTrace, mixed at a 1:1 ratio and transferred into 8-week-old Rag2−/− mice. After 96 h, CellTrace dilution analysis was performed; i.v., intravenous. b, Proportions and CellTrace fluorescence of CD45.1+ FLOX and CD45.2+ CKO T cells before and 96 h after transfer. Left, representative flow plots. Top right, bar graphs showing the percentages of FLOX and CKO T cells. Bottom right, bar graphs showing the percentages of FLOX and CKO T cells divided more than once; n = 4; ****P < 0.0001. c, Left, representative plots of CFSE staining and side scatters (SSC) of 72-h, anti-CD3/CD28-stimulated FLOX and CKO T cells. Right, bar graphs showing the percentages of CFSElo FLOX and CKO T cells after 72 h of stimulation; n = 3; ****P < 0.0001. d, Cell cycle analysis by BrdU/7-AAD staining of 48-h, anti-CD3/CD28-stimulated CD4+ and CD8+ T cells. Top, representative flow plots. Bottom, bar graphs showing the distribution of FLOX and CKO T cells across G0–G1, S and G2–M phases, respectively. n = 4; ****P < 0.0001. e, Apoptosis assessment by Annexin V/propidium iodide (PI) staining of FLOX and CKO T cells in the spleen. Top, representative flow plots. Bottom, bar graphs showing the proportions of live (Annexin V–PI–), early apoptotic (Annexin V+PI–) and late apoptotic (Annexin V+PI+) splenic T cells in FLOX and CKO mice; n = 4. Exact P values from left to right: **P = 0.0041, **P = 0.0040, *P = 0.0197, ***P = 0.0004, ***P = 0.0006 and ***P = 0.0009. f, Apoptosis detection performed the same as in e in the lymph nodes. Left, representative flow plots. Right, bar charts showing the proportions of the indicated populations in FLOX and CKO mice (FLOX n = 3, CKO n = 4). Exact P values from left to right: **P = 0.0031, *P = 0.0137, **P = 0.0021, **P = 0.0050, *P = 0.0293 and **P = 0.0053. n refers to the number of biologically independent samples. Error bars represent mean ± s.e.m. (b–f). Data were analyzed by two-tailed, unpaired t-test (b–f).

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