Fig. 1.

EOD extended lifespan and delayed lethal neoplastic disease but had limited effects on aging rate. a Schematic illustration of the experimental design. b Estimated food intake per animal (number of cages at initiation AL: n = 18, EOD: n = 17). c Body weight (AL: n = 72 mice, EOD: n = 68 mice at initiation). Statistical analyses for b and c were performed using a mixed effects model with DR as fixed effect and cage and time as random effects. d Survival curves (log-rank test; AL: n = 43 mice, EOD: n = 37 mice). e Maximum lifespan was calculated from the longest living 20% (t-test; AL: n = 9 mice, EOD: n = 8 mice). f Causes of natural deaths in AL (n = 29 mice) and EOD (n = 20 mice) animals. g Tumor burden in animals that died of natural causes (AL: n = 29 mice, EOD: n = 20 mice). h–j Summary of large-scale phenotyping analyses to study organismal aging in AL and EOD mice. h The Venn diagram provides an overview regarding the total number of phenotypes assessed (n = 239), the number of aging traits (n = 116), and parameters modified by EOD (n = 89), as well as overlaps of these sets of measures. Forty aging traits were ameliorated by EOD, of which 7 were prevented by EOD and 33 were modulated with similar outcomes in young mice, treated for only 1 month, and in aged animals kept on almost lifelong EOD. Fourteen aging traits were exacerbated by EOD. i Effect sizes (Cohen’s d) of age are plotted against effect sizes of EOD (in old mice) for all aging traits (black dots; n = 116), as well as for all traits modulated by both age and dietary intervention (red dots; n = 53). In both cases, the data show an inverse correlation of the effects of age and DR. j Effect sizes (Cohen’s d) of EOD in young mice are plotted against effect sizes of EOD in old mice for all traits modulated by diet (black dots; n = 88), as well as for all aging traits opposed by EOD (red dots; n = 39). In both cases, the data show a correlation of EOD effect sizes in young and old mice. AL ad libitum, EOD every-other-day feeding