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. 2015 Dec 22;26(2):229–238. doi: 10.1038/cr.2015.148

Figure 3.

Figure 3

spr-5(by101) transgenerational longevity is dependent on daf-12 and daf-36. (A) spr-5(by101) mutant worms live longer than wild-type worms at generation 20 but spr-5;daf-12 double mutant worms do not live longer than daf-12(m20) mutant worms (two-way ANOVA P < 0.0001). Statistics are presented in Supplementary information, Table S4. (B) spr-5;daf-12 double mutant worms show similar levels of H3K4me2 as spr-5(by101) mutant worms at generation 20 as assessed by western blots of whole worm lysates of L3 worms. (C) spr-5;daf-12 double mutant worms lay similar numbers of eggs as spr-5(by101) mutant worms at generation 20 (graph is the mean ± SEM of three independent experiments: each experiment consists of average eggs laid for 10 worms of each genotype performed in triplicate). *P < 0.05 by paired t-test; ns, not significant by two-way ANOVA. (D) daf-36 knock down eliminates the spr-5(by101) lifespan extension at generation 15 (2 way ANOVA P < 0.0001). spr-5 EV lifespan extension is compared with WT EV and spr-5 daf-36 lifespan extension is compared with WT daf-36. Statistics are presented in Supplementary information, Table S5. (E) daf-36 knock down does not affect the increased H3K4me2 of spr-5(by101) mutant worms at generation 15 as assessed by western blots of whole worm lysates of L3 worms. (F) daf-36 knock down does not significantly alter the reduced egg laying capacity of generation 15 spr-5(by101) mutant worms. Graph represents the mean ± SEM of 2 independent experiments: each experiment consists of 3 replicates of 10 worms each. ***P < 0.001 by t-test; ns, not significant by two-way ANOVA. (G) Generation 15 spr-5(by101) mutant worms display higher levels of daf-36 mRNA compared with wild-type worms. Graph represents the mean ± SEM of two independent experiments: each experiment consists of 2 biological replicates of 100 worms each. **P < 0.01 by t-test. (H) Dafachronic acid addition extends wild-type worm lifespan (14.5% P < 0.0001) and generation 6 spr-5(by101) mutant worm lifespan (19.6% P = 0.0005). Statistics are presented in Supplementary information, Table S5. (I) Dafachronic acid addition extends wild-type worm lifespan (30.2% P < 0.0001) but does not extend generation 15 spr-5(by101) mutant worm lifespan (3.9% P = 0.2372). Statistics are presented in Supplementary information, Table S5.