Fig. 1: Neuronal guanylate cyclase regulates peripheral fat mobilization.

a SRS microscopy images and quantification of fat content levels in the anterior intestine of 3-day-old adults show increased fat storage in the guanylate cyclase daf-11 mutants, when compared to wild-type worms (WT). Data are mean ± s.e.m., **P < 0.01 by one-way ANOVA with Dunnett’s test, scale bar = 40 μm, dashed lines indicate quantified intestine areas, yellow pixels indicate higher SRS signals. b Triglyceride levels measured using colorimetric enzyme assays are increased in the daf-11 mutants, when compared to WT. Data are mean ± s.d. of five biological replicates with ~5000 worms for each genotype in each replicate, *P < 0.05, ***P < 0.001 by one-way ANOVA with Dunnett’s test. c–e daf-11 mutants do not show any significant changes in their locomotion (c), or in feeding (d) and defecation rates (e) when compared to WT. Statistical analysis with one-way ANOVA with Sidak’s multiple comparison test. The boxes span the interquartile range, median is marked by the line and whiskers indicate the minimum and the maximum measurements. f Labeling/chasing assays with deuterium-labeled oleic acid (OA-D₃₄) determine rates of lipid synthesis and catabolism. Signals derived from deuterium-labeled lipids in intestinal lipid droplets were quantified at indicated time points using SRS microscopy. daf-11 mutants have no changes in the rate of lipid synthesis, but have a significant reduction in the rate of lipid catabolism (P < 0.01 by linear regression analysis). Data are mean ± s.d, dashed lines represent the trendlines and their corresponding equations are indicated. SRS intensity curve represents replicate #1 (Replicate #2 is presented in Supplementary Fig. 2c). g Reduction in oxygen consumption rate is measured after addition of etomoxir, that blocks mitochondrial β-oxidation, using Seahorse extracellular flux analyzer. daf-11 mutants have reduced levels of mitochondrial β-oxidation compared to wild-type worms. Data are mean ± s.e.m. of three independent biological replicates with at least ten microplate wells containing about 25 worms, *P < 0.05 by Student’s two-tailed t-test. For a and c–f numbers of animals used are listed in Supplementary Data 1. For a–g source data are provided as a Source Data file.