"we investigated whether GE can increase nuclear DAF-16. As shown in Fig. 5A,
daf-2 RNAi highly induced DAF-16 localization to nucleus. However, GE treatment did not show significant effect (Fig. 5B). To further confirm the induction of
daf-16 expression, we performed Q-PCR of
daf-16 and two of its target genes:
sod-3 and
hsp-16.1 (Fig. 5C-E) [42]. We found that GE treatment significantly up-regulates these three genes in C. elegans, confirming that the lifespan-promoting effect of GE is mediated by the DAF-16 pathway (Fig. 5C-E)."