Fig. 6: Cyrene extends lifespan in daf-16 mutants and modulates DAF-16–associated transcriptional networks.

The role of the FOXO transcription factor DAF-16 in the ability of Cyrene to extend lifespan was examined by measuring the ability of Cyrene to increase lifespan in daf-16 deletion mutants. A, B Treatment with 1% Cyrene significantly extends lifespan in daf-16 mutants indicating that the lifespan-extending activity of Cyrene is at least partially independent of DAF-16. At the same time, deletion of daf-16 decreases the lifespan of Cyrene treated worms. C Overlap and enrichment analysis of genes differentially expressed in Day 8 adults treated with Cyrene from L1, showing that both upregulated and downregulated genes significantly intersect with DAF-16 stress-response targets, consistent with bidirectional modulation. D Heatmap displaying relative expression levels of top Class I (DAF-16–activated) and Class II (DAF-16–repressed) genes, indicating simultaneous up- and downregulation within DAF-16–regulated transcriptional modules. Overall, treatment with Cyrene does not result in any consistent, directional modulation of DAF-16 target genes. Error bars represent the standard error of the mean (SEM). Sample sizes (n-values) are indicated within each bar. Statistical analyses were performed using two-way ANOVA with Šidák’s multiple comparisons test in (A), and log-rank test in (B). ****p < 0.0001 from control.