Fig. 1: HLH-30 protects against a senescence-like state, enabling dormancy and stem cell longevity.
a, ARD lifespan of hlh-30(tm1978) and N2 WT animals. b, Brood size of self-fertilizing worms refed after 48 h of ARD. Genotypes N2, hlh-30(tm1978). Each circle represents the total progeny per worm. One representative experiment. Mann–Whitney test (two sided). Biological replicates (BR) = 3. c, Representative photomicrographs of germ cell nucleoli of N2, hlh-30(tm1978) at 96 h of ARD. Dashed circles denote germ cell nucleoli; arrowhead indicates a DTC. Scale bars, 10 μm. d, Distal gonad arms dissected from worms, refed at 48 h of ARD for 24 h, stained with DAPI (DNA, turquoise) and anti-phospho-histone H3 (M-phase chromosomes, magenta, arrows). Representative photomicrographs. Genotypes N2, hlh-30(tm1978). Scale bars, 10 μm. Arrowhead indicates a DTC. e, Photomicrographs of distal gonad arms of sygl-1::3xFlag worms in N2 and hlh-30(tm1978) backgrounds, stained with DAPI (nucleus, gray), anti-RAD-51 (DNA damage foci, magenta) and anti-3xFLAG (SYGL-1, GSC zone, yellow) antibodies. Following 48 h of ARD, worms were dissected after 24 h of refeeding. Scale bars, 10 μm. f, Three-dimensional (3D) quantification of mitochondrial sphericity in GSCs at 48 h of ARD and 48 h of refeeding. Genotypes N2, hlh-30(tm1978). Each dot represents the mitochondrial sphericity of one individual worm. Pooled data of two independent biological replicates. Two-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. g, Representative images of mitochondrially produced ROS using CM-H2TMROS staining after 48 h in ARD. Scale bars, 10 μm. h, Representative images of SA-β-gal staining after 48 h of ARD. Dashed circles show the two pharyngeal bulbs. Scale bars, 10 μm. i,j, Pharyngeal pumping (i) and body bending (j) rates of 48-h ARD worms. 30-s intervals. Each dot represents the pumping rate or body bends of one animal. Mann–Whitney test. BR = 3, one representative experiment. k, Percentage reproductive worms refed after 10 days of ARD. The genotypes are shown in Table 1. Each circle represents the percentage from one biological repeat. Mann–Whitney test (two sided). Survival curves depict one experiment. All data and statistics are presented in Supplementary Table 2. If not stated otherwise, data are the mean ± s.d. NS, not significant.
