Fig. 6: NAT10 interacts with RACK1 and promotes its acetylation.

a CFSE labeled WT naïve CD4⁺ T cells were activated in complete medium containing 20 µM Remodelin, and the proliferation rate was measured at 48 h and 72 h (n = 3). D, DMSO; R, Remodelin. b Cell cycle analysis of αCD3 + αCD28-activated T cells treated with 20 µM Remodelin was evaluated by BrdU incorporation (n = 3). c Venn plot showing overlapped proteins which both interacted with NAT10 and acetylated in activated T cells. d RACK1 were isolated by IP (under denaturing conditions) from whole-cell lysates of naive CD4⁺ T cells stimulated with αCD3 plus αCD28 and subjected to IB assays using anti-pan acetylation (top panels) antibodies. Protein lysates were also subjected to direct IB (bottom panels). e The activated T cells were fixed and permeabilizated, stained with primary antibodies and fluorescein-conjugated second antibody (Red for RACK1; Green for NAT10; Blue for DAPI), representative images were subsequently captured by confocal. The left scale bar indicates 10 μm, and the right scale bar indicates 2 μm. f Structure (top) and functional domain (bottom) of human NAT10 protein. The different colors indicated distinct domain of NAT10. g RACK1 were isolated by immunoprecipitation (IP) and subjected to IB assays using FLAG (top panels) antibodies to evaluate the interaction between RACK1 and NAT10. Protein lysates were also subjected to direct IB (bottom panels). 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, b). **P < 0.01, ***P < 0.005.