Fig. 1: Different modes of sterility induced excessive fat accumulation but differential lifespan.

a Survival curves showing the lifespans (at 20° C) of wild-type worms treated with the indicated gene RNAi at 20 °C. b, c Survival curves showing the lifespans of wild-type N2, glp-1(e2144), fem-3(e1996), and mog-3(q74) mutants grown at 25 °C, then aged at 20 °C (b), or continuously at 25 °C (c). The detailed mean lifespan values, the number of worms analyzed in each replicate, and statistical analyses for (a–c) are provided in Source Data. Two biological replicates were analyzed for each genotype. d, e Lipid levels estimated by Oil Red O (ORO) staining (d) and triacylglycerides (TAG) quantification by LC-MS, normalized to N2 (e), in the indicated genotypes at 25 °C. ORO intensity is shown as the means of two biological replicates per genotype, with at least 10 worms per replicate and 20 worms total per genotype. In (d), the p-values for comparisons with N2 are 8.93 × 10⁻¹⁴ for glp-1(-) ts; 9.02 × 10⁻¹¹ for fem-3(-), and 2.94 × 10⁻⁹ for mog-3(-), respectively. In (e), the p-values for comparisons with N2 are 0.00014 for glp-1(-) ts; 0.00032 for fem-3(-), and 0.02345 for mog-3(-), respectively. TAG estimations in (e) are based on three biological replicates from the lipidomics LC-MS dataset. f, g Representative images (f) and quantification (g) of VIT-2::GFP intensity in glp-1(RNAi), fem-3(RNAi), and mog-3(RNAi) worms compared to Empty Vector (EV) RNAi controls. VIT-2::GFP intensities are shown as the mean of two biological replicates per genotype, with at least 10 worms per replicate and 20 worms total per genotype, except for glp-1(e2144), for which 19 worms were analyzed. In (g), the p-values for comparisons with empty vector (EV) are 4.24 × 10⁻¹⁹ for glp-1(RNAi); 3.63 × 10⁻¹⁴ for fem-3(RNAi), and 5.72 × 10⁻¹³ for mog-3(RNAi), respectively. In (d, e, and g), unpaired two-tailed Welch’s t-tests were used to perform one-way comparisons of each genotype to empty vector (EV) RNAi control. No correction for multiple comparisons was applied. p-values for the tests are indicated as *p < 0.05, ***p < 0.001, ****p < 0.0001. Error bars represent standard deviations. Scale bar in (f), 100 µm. LC-MS - Liquid Chromatography–Mass Spectrometry.