Figure 3. Lifespan extension induced by thymine treatment is mediated by the nuclear receptors DAF-12 and NHR-49, and the transcription factor DAF-16/FOXO. (A) Effects of thymine supplementation at increasing concentrations (0.05-10 mM) (P value determined by log-rank tests). (B) Age-related movements of worms treated with 2 mM thymine and vehicles. Data are the mean ± SD; * P <0.05 (Student’s t test). (C–F) Lifespan analysis in (C) daf-12 (rh274), (D) daf-16 (mu86), (E) nhr-49 (gk405), and (F) skn-1 (zu67) worms treated with 2 mM thymine (red) and vehicle (black), on heat-inactivated E. coli OP50 (P value by log-rank test). Lifespan values of replicated experiments are summarized in Supplementary Table 1. (G) QPCR analysis of the mRNA level of target genes of daf-12, daf-16 and nhr-49 in C. elegans treated with 2 mM thymine versus control. Data are the mean ± SD, n=3, * p<0.05 (Student’s t test). (H–I) Images and quantification of GFP fluorescence. *p < 0.05; n=30 (Student’s t test).