Fig. 4: Pol I inhibition decelerates metabolic aging.

a Survival analysis of C. elegans wild-type (N2 Bristol) and ife-2 mutant strains treated with control (Ctrl) RNAi or tif-1A RNAi, n = 140 worms. b Comparison of ATP pools at AD2 between wild-type (N2) and ife-2 mutant worms treated with Ctrl RNAi or tif-1A RNAi. ATP levels were measured in a bioluminescence assay and normalized to total protein content. Values are shown relative to N2 Ctrl RNAi. Data are from 6 independent experiments with n = 50 worms in each sample. Mean ± S.D. values are shown. c Comparison of ATP pools at AD2 and AD12 between worms treated with Ctrl RNAi or tif-1A RNAi. ATP measurement was conducted as in b, and values are relative to Ctrl RNAi on AD12. Mean ± S.D. values of 6 independent experiments with n = 50 worms are shown. d Survival analysis of worms exposed to Ctrl RNAi or tif-1A RNAi and, from AD12 onwards, to 50 mM metformin (Met), n = 140 animals. A representative result of four independent experiments is shown. For a and d, survival was scored daily and P values were determined using the Mantel-Cox test. For b and c, a Student’s unpaired t-test was used for statistics. e, f BJ primary human skin fibroblasts were treated with indicated doses of metformin in the absence (DMSO) or presence of the indicated concentrations of ATP (positive survival control) or the Pol I inhibitors BMH-21 (e) and CX-5461 (f). Mitochondrial activity was assessed by MTT assay, and an unpaired Student’s t-test was used to compute P values, comparing each co-treatment to the respective DMSO + Metformin control condition, data represent mean ± S.D of 3 independent experiments. ns = not significant, ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. All statistical tests used were two-sided, the exact P values and statistical analyses are reported in the Source Data file.