Extended Data Fig. 7. Polyamine synthesis is important for IF-induced benefits in flies and worms.

(a) Daily and nightly food consumption of D. melanogaster w¹¹¹⁸ after 10, 30 and 50 days of IF^12:12. N = 11(Day 30 IF Day), 12(Day 30 Ad lib), 16(Day 50 Ad lib Night), 18(Day 10), 20(Day 50 IF Day), 22(Day 50 Ad lib Day) biologically independent samples (groups of flies). (b) DFMO feeding affects the polyamine profile of female w¹¹¹⁸ flies during ad lib and 24 hours fasting. N = 6(Ad lib 10 µM DFMO; Fasted 0, 10 µM DFMO), 7(Ad lib 0, 0.1, 1 µM DFMO; Fasted 0.1, 1 µM DFMO) biologically independent samples (groups of flies). (c) Lifespan of male w¹¹¹⁸ flies fed standard food with or without 10 mM DFMO and subjected to IF. N = 210(Ad lib), 206(IF), 199(Ad lib+DFMO), 212(IF + DFMO) flies. (d) Food consumption of 10-day old female and male w¹¹¹⁸ flies, fed control or food containing 10 mM DFMO, during the first 7 cycles of IF. N = 8(0 mM Day IF), 9(rest) biologically independent samples (groups of 5 flies per N). (e) DFMO feeding does not influence body weight changes after 24 hours fasting. N = 7 biologically independent samples (groups of flies). (f) Lifespan of heterozygous Odc1^MI10996/+ flies subjected to IF. The IF + SPD group received 5 mM SPD via agar during the night. N = 119(Odc1^MI10996/+ ad lib), 117(Odc1^MI10996/+ IF), 113(Odc1^MI10996/+ IF + SPD) worms. (g) Knockdown of odc-1 does not affect worm size under ad lib or IF^48:48 conditions, compared to CTL RNAi. Note that the experiment and statistics were performed together with the dhps-1(RNAi) groups in Extended Data Fig. 10h. N = 56(CTL ad lib), 66(CTL IF), 63(Odc-1(RNAi) ad lib), 78(Odc-1(RNAi) IF) worms. (h) Knockdown efficiency of respective mRNAs by feeding bacteria expressing argn-1, spds-1, or smd-1 RNAi in C. elegans. N = 2(argn-1, smd-1), 3(spds-1) biologically independent experiments. (i) Lifespan of C. elegans N2 fed control (CTL) or RNAi against odc-1, with or without continuous 0.2 mM SPD feeding during IF. Note that the experiments and statistics were performed together with the groups depicted in [J]. N = 429(CTL ad lib), 523(CTL IF), 437(CTL ad lib+SPD),466(IF + SPD), 330(odc-1(RNAi) ad lib), 575(odc-1(RNAi) IF), 497(odc-1(RNAi) ad lib+SPD), 605(odc-1(RNAi) IF + SPD) worms. (j) Lifespan under 50 µM rapamycin treatment of C. elegans N2 fed control (CTL) or RNAi against odc-1. Note that the experiments and statistics were performed together with the groups depicted in [I]. N = 429(CTL), 423(Rapa), 330(odc-1(RNAi), 330(odc-1(RNAi) Rapa) worms. Statistics: [A,D] Two-tailed Student’s t-test (per time point). [B,E,G] Two-way ANOVA with Holm-Šídák’s multiple comparisons test. [F,I,J] Log-rank test with Bonferroni correction. [H] Two-tailed Student’s t-test. Heatmaps show means. Bar graphs show the mean ± s.e.m. Source numerical data are available in source data.

Source data