Figure 3. DAF-2B influences dauer entry in temperature-sensitive insulin-signaling mutants.

(A) DAF-2B overexpression (OE) from the native daf-2 promoter, in the nervous system or in the hypodermis enhances dauer formation in daf-2 mutants at 23.2°C. There was no effect of DAF-2B expressed in the intestine or in the body wall muscle. Data are pooled from 2 (hypodermal promoter) or 3 (all other promoters) transgenic lines with at least four biological replicates per transgene. Students t-test ns – not significant, **p<0.01, ***p<0.001. Two additional trials with native, neuronal, muscle and intestinal DAF-2B OE showed similar effects. Raw data can be found in Figure 3—source data 1. (B) Schematic illustrating the region of the daf-2 genomic locus deleted to generate daf-2bc(Δ). (C) Genetic deletion of daf-2b suppresses dauer formation in pdk-1 mutants at 26.8°C. Data are pooled from three independent trials with six biological replicates from one control and one daf-2b(Δ) strain per trial. Students t-test ***p<0.0001. Raw data can be found in Figure 3—source data 2. (D) Genetic deletion of daf-2c has no effect on dauer formation in pdk-1 mutants at 26.8°C. Data are pooled from three independent trials with three biological replicates from each of two control and daf-2c(Δ) strains per trial. Students t-test ns – not significant. Raw data can be found in Figure 3—source data 3. (E) In temperature-sensitive hypomorphic insulin signaling mutants, the decision to enter dauer or develop into reproductive adults is dependent on the level of insulin agonism. (F) Under semi-permissive conditions, agonism at the insulin receptor promotes insulin signaling and reproductive growth. (G) DAF-2B, acting as a homodimer or a heterodimer, is predicted to sequester agonist insulin peptides, thereby inhibiting insulin signaling and promoting dauer formation. Error bars represent mean ± sd and data are shown as independent replicates from multiple transgenic lines.