Extended Data Fig. 5: NAT10 knockdown compromised proliferative potency of P14 T cells in response to LCMV infection.
a Schematic diagram of in vivo experiments to explore the role of NAT10 in specific-antiviral response of P14 T cells. CD45.1+ CD8+ P14 T cells were isolated, labeled with CFSE, and then infected by retrovirus containing LMP-NC and LMP-Nat10-shRNA plasmids, respectively. Equal amounts of these T cells (4 × 105 cells) were then i.v. injected into 8-week-old CD45.2+ recipient mice 1 day before Armstrong infection. 3 or 7 days later, the recipients were sacrificed for proliferation analysis. b Knockdown efficiency of NAT10 was assessed by western blot analysis. Representative data of three independent experiments is presented. c CFSE dilution analysis before injection and 72 h after LCMV infection. Left, representative flow plots. Right, bar graphs showing percentages of CFSElo P14 T cells in the 2 groups, respectively. n = 4, ****P < 0.0001. d Proportion of retrovirally infected P14 T cells identified by AmCyan fluorescence. Left, representative flow plots. Right, bar diagrams showing the percentage, absolute number and expansion folds of AmCyan+ P14 T cells in each group. n = 4, ****P < 0.0001, ***P (#AmCyan+ P14 T cell) = 0.0003, ***P (Expansion foldchange) = 0.0003. e Detection of IFNγ+ TNF+ AmCyan+ P14 T cells in recipients with control or NAT10 KD P14 T cells adaptive transfer, respectively. Up, representative flow plots. Down, bar diagrams exhibiting percentages and absolute number of splenic IFNγ+ TNF+ AmCyan+ P14 T cells in the two groups. n = 4. ****P < 0.0001. The values of “n” all refer to biologically independent samples. Error bars represent mean ± s.e.m. (c–e). Two-tailed, unpaired t-test (c–e).
