Fig. 3

Ethosuximide-induced genes are enriched in DAF-16-associated elements and ethosuximide-induced lifespan extension requires daf-16. a Validation of gene expression changes using qRT-PCR. Selected genes that were up-regulated by ethosuximide in microarray experiments (ugt-25, dhs-26, cyp-14A3, cyp-35B1, ttr-44, dod-6 and cyp-34A9) were confirmed to be significantly induced using qRT-PCR. No significant changes in expression of normalisation (act-1, pmp-3) or negative control (pph-6) genes was observed. Results are expressed as mean fold change ± SEM relative to the unexposed control (n = 3). b Ethosuximide-responsive genes are enriched for the DAF-16 Associated Element (DAE) motif. To identify regulatory sequences correlating with ethosuximide-responsiveness, 200-bp regions in the upstream promoter sequences of common DEGs were mined for overrepresented motifs using RSAT. c Ethosuximide increases dnj-14 lifespan in a daf-16-dependent manner. Survival curves of dnj-14(ok237) worms grown on E. coli containing empty vector (L4440), hsp-1 or daf-16 dsRNA-producing plasmids in the presence (dashed lines) or absence (solid lines) of 1 mg/ml ethosuximide. Ethosuximide treatment significantly increased the lifespan of dnj-14(ok237) worms on vector control (p < 0.001), but had no significant effect on daf-16 RNAi animals (p > 0.15) (n > 100 worms per strain/condition)