Fig. 3

NAD⁺ replenishment ameliorates accelerated aging in WS worms and flies. a Relative NAD⁺ levels in adult D2 worms. Data are shown in mean ± S.E.M (n = 3 biologically independent experiments; Two-way ANOVA). b Lifespan curves in N2 and wrn-1(gk99) worms at 25 °C. c Effects of NR/NMN on lifespan in wrn-1(gk99) worms at 25 °C. Experiments were repeated seven times. One representative set of data is shown. Summarized data, including effects of NR/NMN on N2 worms, are in Supplementary Table 4. d Effects of NR/NMN on pharyngeal pumping in N2 and wrn-1(gk99) worms. Data, mean ± S.E.M (n = 3 biologically independent experiments with 10–20 worms for each condition; Two-way ANOVA). e Changes of germ line-localized mitotic cells in designated groups of worms. Data, mean ± S.E.M (n = 10 worms per condition; Two-way ANOVA). f Differences of the numbers of pH3⁺ mitotic cells in the designated groups of worms after exposure to 90 Gy ionizing radiation. Data, mean ± S.E.M (n = 6 worms per condition; Two-way ANOVA). g Effects of NAD⁺ supplementation on lifespan in w;DaGeneSwitch-Gal4;y,v,sc;Wrnexo^RNAi (EtOH) (WT), and w;DaGeneSwitch-Gal4;y,v,sc;Wrnexo^RNAi (RU486) (WRN-KD) flies at 25 °C without/with 5 mM NMN treatment. A representative set of data from two biological repeats is shown. Log-rank test was used for statistics: ***p < 0.0001 compared with WT (EtOH), ^###p < 0.0001 compared with WRN-KD (EtOH). h, i Effects of NMN (h) and NR (i) on gut damage-induced proliferating intestinal stem cells (pH3⁺, ISCs) in adult D7 flies. (n = 10 flies for each condition, two biological repeats). One-way ANOVA using Sidak’s post-hoc test. Data are shown in mean ± S.E.M. *p < 0.05, **p < 0.01, ***p < 0.001.