Lifespan analysis

All lifespans were done at 20 °C. Synchronised eggs were obtained by hypochlorite treatment of adult hermaphrodites and placed on NGM plates containing OP50 E. coli bacteria. During the course of the lifespan, we transferred wild-type adult nematodes every day during their reproductive period and afterwards on alternate days. We separated homozygous phb-1(tm2571) and phb-2(tm2998) mutants from their respective heterozygous populations at L3 stage to a separate plate and transferred them every alternate day. Worms were scored as dead when they stopped responding to prodding, while exploded animals, those exhibiting bagging, protruding gonad or drying out on the edge of the plates were censored. We used GraphPad Prism software to plot survival curves, and we determined significant differences in lifespan by using the log-rank (Mantel-Cox) test.