Fig. 3

AAK-2 regulates the catabolism, the gluconeogenic mode, and the developmental arrest in the dauer state. a Electron microscopy of daf-2(e1370);aak-2(gt33) collected on the day of the dauer formation or after 5 days of incubation at 25 °C on ample food source. Alae: arrows. Striated layer of the cuticle: brackets. Mitochondria: arrowheads. Starvation features: shrinkage of the hypodermis (H), expansion of the pseudocoelomic cavity (1 asterisk), a cavity between the cuticle and the hypodermis (2 asterisks), widening of the gut lumen (3 asterisks). Scale bars 5 μm (upper panels) and 0.5 μm (lower panels). Representative images of at least 4 animals. b TLC of ¹⁴C-acetate-labeled lipids and sugars from daf-2 and daf-2;aak-2 dauers measured at different time points after the arrest. TG, triglycerides; GlcCer, glucosylceramides; Mar, maradolipids; PE, phosphatidylethanolamines; PS, phosphatidylserines; PI, phosphatidylinositols; PC, phosphatidylcholines; Tre, trehalose. c Quantification of band intensities in some of the compounds in b represented by the peak area of the optical density in relative units (RU). Means ± SD of 2 experiments with 3 biological replicates. ***p < 0.001; **p < 0.01; *p < 0.1; ns, no significant difference determined by Student t test. d 2D-TLC of ¹⁴C-acetate-labeled metabolites from daf-2;aak-2 L3 larvae grown at 15 °C, dauers grown at 25 °C, and L3 larvae obtained at 25 °C in the presence of DA. DA, dafachronic acid; Tre, trehalose; Glc, glucose; Glu, glutamate; Gly, glycine; Ser, serine; Gln, glutamine; Ala, alanine; Thr, threonine. Representative images from at least 2 experiments. e Micrographs of daf-2 and daf-2;aak-2 animals grown at 25 °C in the presence or absence of DA. f Quantification of the larval arrest in e. Means ± 95% confidence intervals of 3 experiments with 3 replicates. ***p < 0.001; ns, no significant difference determined by one-way analysis of variance