Fig. 10: efk-1 mutants exhibit mitochondrial defects in starvation.

A The graph shows L1 starvation survival of WT and efk-1 mutants with or without supplementation of mitochondrial antioxidant MitoQ at 5 µM. N = 4, error bars represent mean ± SD; ***p = 0.0006, ****p < 0.0001 (AUC compared using one-way ANOVA with Tukey’s multiple comparisons test). B, C Assessment of muscle mitochondria morphology in WT and two lines bearing an identical ok3609 mutation, efk-1(ste4) and efk-1(ste5), referred to as efk-1 line 1 and line 2, respectively. Synchronized L1 worms bearing myo-3::GFP(mit) were imaged post hatching, and after 6 and 9 days of starvation. The percentage of worms showing complete fragmentation is quantified in C. N = 3 (total 30 worms per condition), error bars represent mean ± SD; **p = 0.0028 efk-1 line 1, **p = 0.0052 efk-1 line 2 at day 6, *p = 0.0334 efk-1 line 1, *p = 0.0334 efk-1 line 2 at day 9 vs. respective WT controls; ^##p = 0.0015 WT day 9, ^##p = 0.0052 efk-1 line 1 day 6, ^###p = 0.0002 efk-1 line 1 day 9, ^###p = 0.0008 efk-1 line 2 day 6, ^####p < 0.0001 efk-1 line 2 day 9 vs. respective day 0 controls (two-way ANOVA with Dunnett’s multiple comparison’s test). Scale bar, 10 µm. D The graph shows continual oxygen consumption rate (normalized OCR per worm, y-axis) of WT and efk-1 mutants with or without the rescue construct efk-1::GFP in L1 starvation (hours of starvation, x-axis). N = 3-4, shaded area represents mean ± standard error of the mean (SEM); *p = 0.0219, ****p < 0.0001 vs. WT, ^####p < 0.0001 vs. efk-1 (one-way ANOVA with Tukey’s multiple comparisons test). E Model of how efk-1 promotes starvation survival via a noncanonical pathway. Created in BioRender. Yan, J. (2025) https://BioRender.com/d17w564. WT, wild type; ns, not significant; ROS, reactive oxygen species; NAC, N-acetylcysteine; AUC, area under the curve; MitoQ, mitoquinone; OCR, oxygen consumption rate. Source data are provided in the Source Data file and Supplementary Data 4.