C. elegans Lifespan Imaging Protocol

C. elegans strains were maintained on nematode growth media (NGM) plates with Escherichia coli OP50-1 bacteria at 20°C as described by Brenner (1974) (link). SK4005: zdIs5 [Pmec-4::GFP + lin-15(+)] was used and is referred here as WT control. The daf-2(e1370) mutation was crossed into SK4005. Strains used in the experiments were maintained for at least two generations at 20°C. Synchronization was done by hypochlorite solution with direct transfer of the bleached eggs to NGM plates. C. elegans were grown to L4 at 20°C and transferred to 50 μM 5-fluorodeoxyuridine (FUDR; Sigma-Aldrich F0503) plate to be maintained at 25°C (Teuscher et al., 2019 (link)). At day 1 of adulthood, ∼20 animals per condition were used for imaging, the rest were maintained to day-8 adulthood at 25°C for the second imaging session. For the experiments at 15°C, all processes are the same as above except that all C. elegans were always kept at 15°C. RNAi was performed as described in Ewald et al. (2017) (link).