Fig. 2: NAT10 helps to maintain the pool of T cells.

a, NAT10 protein expression in CD3⁺ T_N cells stimulated with anti-CD3/CD28 for the indicated lengths of time (top) or CD4⁺ and CD8⁺ T_N cells stimulated for 24 h from FLOX and CKO mice (bottom). b, Dot blot and densitometry analysis of ac⁴C modifications in total RNA (1,000 ng) from CD4⁺ and CD8⁺ T_N cells stimulated with anti-CD3/CD28 for 24 h. Left, representative anti-ac⁴C dot blot. Right, bar graph showing the relative ac⁴C intensities quantified by methylene blue; n = 3; **P = 0.0037 and *P = 0.0103. c, Comprehensive t-SNE visualization of splenic and thymic single cells from 6- to 8-week-old FLOX and CKO mice by cell type; NK, natural killer. d, Separate displays of c by different group, with the same color coding. e, Bar plot showing the percentages of each cluster across groups; related to d; S, splenic; T, thymic. f, Representative FCM plots of splenic CD3⁺TCRβ⁺, CD4⁺ and CD8⁺ T cells in 8-week-old FLOX and CKO mice in vivo. g, Percentage of CD3⁺TCRβ⁺ T cells and absolute numbers of CD3⁺TCRβ⁺, CD4⁺, CD8⁺ and double-negative (DN) T cells in the spleens of FLOX and CKO mice; n = 3; ****P < 0.0001, ***\({P}_{{\rm{No.}}\, {\rm{CD}}3^{+}{\rm{TCR}}\upbeta^{+}}\) = 0.0010, ***\(P_{\rm{No}. \rm{CD8}^+}\) = 0.0003, ***P_{No. DN} = 0.0008 and **P = 0.0031. h, Representative FCM plots of CD3⁺TCRβ⁺, CD4⁺ and CD8⁺ T cells in inguinal lymph nodes of FLOX and CKO mice. i, Percentage of CD3⁺TCRβ⁺ T cells and absolute number of CD3⁺TCRβ⁺, CD4⁺, CD8⁺ and double-negative T cells in inguinal lymph nodes of FLOX and CKO mice; n = 3; ****P < 0.0001, ***P = 0.0001, **P = 0.0022 and *P = 0.0183. j, Proportions of CD3⁺TCRβ⁺ T cells in spleen, lymph node, lung, kidney and liver of FLOX and CKO mice; n = 3. ****P < 0.0001, **P_{Lymph node} = 0.0081, **P_Lung = 0.0022 and *P = 0.0104; NS (not significant), P > 0.05. k, Thymic double-negative, double-positive (DP) and CD4⁺ and CD8⁺ single-positive (SP) T cells in FLOX and CKO mice. Left, representative FCM plots. Right, bar graphs showing proportions and absolute numbers of indicated thymic populations in FLOX and CKO mice; n = 3. n refers to the number of biologically independent samples. Error bars represent mean ± s.e.m. (b, g and i–k). Representative data of three independent experiments are presented (a). Data were analyzed by two-tailed, unpaired t-test (b, g and i–k).

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