Fig. 3: ATF-4 overexpression is sufficient to increase lifespan.
a Transgenic animals (wbmEx26 [Patf-4::ATF-4(gDNA)::GFP]) that overexpress ATF-4 (ATF-4OE) live longer compared to their non-transgenic siblings. b Pharyngeal pumping rate is similar at day 2 of adulthood between ATF-4OE (ldIs119 [Patf-4::ATF-4(gDNA)::GFP]) and WT, but higher in ATF-4OE at day 10 of adulthood, suggesting an improved healthspan. For the complete time-course of pharyngeal pumping rate during ageing, see Supplementary Data 2. Mean ± SEM. At least 5 worms were examined for each genotype and time point in one experiment. Unpaired two-tailed t-test. c MA (log ratio and mean average)-plot of RNA sequencing analysis comparing ATF-4OE(ldIs119) to WT, showing absolute log fold change (FC). In red, highlighted genes with FDR < 0.1 and log FC > 1 compared to WT. In black, genes with FDR > 0.1. Details are in Supplementary Data 3. d Validation by qRT-PCR of genes differentially expressed in ATF-4OE(ldIs119), using two new independent biological samples of about 200-215 animals each measured in duplicates. Mean ± SEM. P values relative to WT determined by one sample t-test, two-tailed, hypothetical mean of 1. The numbers of ATF4 binding sequences (-TGATG-)27,28 are indicated in Supplementary Data 4. The DAF-16 and SKN-1 transcription factor binding sites are based on ChIP data from www.modencode.org (Supplementary Data 5). e Longevity conferred by ATF-4OE(ldIs119) is abolished by knockdown of hsf-1, skn-1, or daf-16. Mean ± SEM of Kaplan–Meier survival plot. P-value determined by log-rank. n = 1 biological replicate. f The mitochondrial ATP translocase ant-1.3 is required for ATF-4 overexpression induced longevity. For statistical details and additional lifespan trials in (a), (e), and (f), see Supplementary Data 1.
