(A) Starvation survival curves are plotted for wild-type (WT), tps-1, tps-2, tps-1; tps-2, and daf-16. (B and C,E and F) Representative Ptps-1::GFP (BC14885) and Ptps-2::GFP (BC14876) worms either fed or starved for ~12 hr are shown with false coloring to depict relative fluorescent intensity according to the calibration bar. Scale bars are 50 μm. (D and G) Boxplots indicate the distribution of average pixel intensity per worm in each reporter background in fed and starved worms. Each point represents an individual worm sampled from one of three biological replicates. Ptps-1::GFP was significantly brighter during starvation (p=0.04, unpaired t-test, n = 3). Ptps-2::GFP intensity was not different between fed and starved worms (p=0.50, unpaired t-test, n = 3). (H) Trehalose levels of WT, tps-1, tps-2, and tps-1; tps-2 are plotted. tps-1 and tps-2 each had reduced levels of trehalose compared to WT (p=0.05, p=0.01, unpaired t-test, n = 4). The double mutant was significantly different from WT and each single mutant (p=0.001, p=0.01, p=0.03, unpaired t-test, n = 4). (I) Median survival time is plotted for a worms exposed to a range of concentrations of trehalose in the buffer in which worms are starved. The dashed line indicates median survival of control animals. (J) Trehalose content 1 d after hatching is plotted for worms exposed to a range of concentrations of trehalose in S-basal (n = 12). The dashed line indicates trehalose content of control animals. (K) L1 larval size is plotted for WT worms with and without trehalose supplementation. Worms starved for 8 d are shorter than worms starved for 1 d (p=1.3E-05, unpaired t-test, n = 5). Worms supplemented with trehalose remain larger after 8 d of starvation (pint=0.007, two-way ANOVA, n = 5). (L) Starvation survival curves are plotted for WT, tps-1; tps-2 (four biological replicates), daf-2, and tps-1; tps-2; daf-2 (three biological replicates). There is a significant interaction by two-way ANOVA between median survivals (pint = 0.02). (M) Starvation survival curves for WT and daf-16 with and without 50 mM trehalose supplementation. (N) Trehalose content is plotted for N2 and daf-16 with and without 50 mM trehalose supplementation (n = 7). Mean and standard error of the mean of biological replicates are plotted in H–K and N.