FIGURE 3 from Urolithin-A Promotes CD8⁺ T Cell–mediated Cancer Immunosurveillance via FOXO1 Activation

UroA induces FOXO1 transcriptional activity in CD8⁺ T cell. A, Quantification of FOXO1 target genes in CD8⁺ T cells culture with 5 µmol/L UroA in IL15/7 condition in vitro. Values are expressed as fold change compare with control condition. B, Analysis of FOXO1 target genes expression levels in CD8⁺ T cells isolated from mice fed with control or UroA-enriched food for 4 weeks. Values are expressed as fold change compare with control condition. C, FOXO1 phosphorylation in CD8⁺ T cells treated for 24 hours with 5 µmol/L UroA in vitro. D, FOXO1 phosphorylation in CD8⁺ T isolated from mice fed with control or UroA diet. E, Representative confocal images of CD8⁺ T cells stained with FOXO1 (green) CD8 (red), and DAPI (blue) under the indicated culture condition (scale bar, 5 µm) in vitro (UroA = 5 µmol/L). The LUT images represent FOXO1 intensity staining (left). Quantification of nuclear FOXO1 in control and UroA (5 µmol/L) treated CD8⁺ T cells) in vitro (right). F, Akt S473 phosphorylation in CD8⁺ T cells treated for 24 hours with 5 µmol/L UroA in vitro and then stimulated with CD3/CD28 antibodies for 30 minutes (Act condition). Data are mean ± SEM. Each dot represents a biological replicate. In A, sample size n = 6. In B, sample size n = 5. In C, sample size n = 8. In D, sample size n = 5. In E, control = 72, UroA = 68. In F, sample size n = 4. Data were analyzed by two-sided Student t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). Representative results of at least two independent experiments or two pooled experiments.