Fig. 4: NAT10 tightly prompts T cells to enter the cell cycle and proliferation.

Naïve CD4⁺ T cell (CD44^loCD62L^hi) derived from NAT10^WT and NAT10^cKO mice were labeled with CFSE and stimulated with anti-CD3 plus anti-CD28 (a) or PMA plus ionomycin (b) for indicated time (n = 3). The proliferation potential was assessed according CFSE dilution by FACS. c Naïve CD4⁺ T cells from NAT10^WT and NAT10^cKO mice were polarized under indicated differentiation condition (n = 3). The subset-specific cytokines or transcription factors were analyzed by FACS after 96 h. d WT and NAT10-deficient naïve CD4⁺ T cell was stimulated with anti-CD3 plus anti-CD28 for 24 h (n = 3). The apoptosis (Annexin V⁺PI⁺) of each group was measured by FACS. e The enrichment in significant terms of TOP20 downregulated genes in NAT10-deficient CD4⁺ T cells compared to those in WT control was analyzed by KEGG analysis. f WT and NAT10-deficient naïve CD4⁺ T cells were stimulated with anti-CD3 plus anti-CD28 for indicated time and incorporated with BrdU (n = 3). The cell cycle was analyzed by FACS. g Heatmap showing different expression level of cell cycle associated genes between NAT10^WT and NAT10-deficient naïve CD4⁺ T cells stimulated with anti-CD3 plus anti-CD28 for 24 h. h Naïve CD4⁺ T cell derived from NAT10^WT and NAT10^cKO mice were stimulated with anti-CD3 plus anti-CD28 for 24 h (n = 3). The mRNA level of Ccne1, Cdk4, Cdkn1a, and Trp53 gene were measured by RT-qPCR. These qPCR data are presented as fold change relative to the Actb mRNA level and normalized by Bio-Rad CFX Manager 3.1. i The protein abundance of c-Myc and P53 in WT and NAT10-deficient CD4⁺ T cells under stimulation were analyzed by immunoblotting (IB). Source data are provided as a Source Data file. Data were represented as mean ± S.D. Three independent experiments at least were performed in data represented above. Statistical significance was determined by unpaired two-tailed t test (a–d, f, h). *P < 0.05, **P < 0.01, ***P < 0.001.