Manipulation of <i>dilp2</i> in IPCs affects HAB in sated state.

Abstract

(A) HAB was not affected in dilp3 mutant flies (dilp3-/-) and dilp5 mutant flies (dilp5-/-) (Satiety: P-values: 0.6672, 0.4866, 0.8345, and 0.4314 from left to right; Hunger: P-values: 0.1133, 0.2915, 0.718, and 0.426 from left to right). (B) dilp2-GAL4 > UAS-dilp2RNAi flies exhibited an increased HAB during the sated state (Satiety: P-values: P-values: 0.6612, 0.7088, 0.2353, and 0.8943 from left to right). (C) Permissive temperature control of Fig 3C. At permissive temperatures, there were no significant differences in HAB between dilp2-GAL4; tub-GAL80ts > UAS-dilp2RNAi, dilp2-GAL4; tub-GAL80ts > +, and UAS-dilp2RNAi > + flies during the sated state (Satiety: P-values: 0.8918, 0.7176, 0.5073, and 0.7762 from left to right). (D) Immunostaining with anti-Dilp2 antibody in dilp2-GAL4 > UAS-mCD8::GFP flies (left panel). Quantification of anti-Dilp2 antibody immunostaining intensity in GFP positive IPCs during sated and hungry states (right panel). The anti-Dilp2 immunostaining signals in IPCs were normalized to the signals in fan-shaped body (P E) Hot stimulus did not induce the calcium response in IPCs during both feeding states. The soma of IPCs were recorded and analyzed. There were no significant differences in GCaMP intensity in IPCs before and after hot stimuli (P-values: 0.8381). The arrow under calcium response curve indicates the time points at which the hot stimuli were applied. (F) Permissive temperature control of Fig 3E. At permissive temperatures, there were no significant differences in HAB between dilp2-GAL4; tub-GAL80ts > UAS-dilp2, dilp2-GAL4; tub-GAL80ts > +, and UAS-dilp2 > + flies during sated and hungry states (Satiety: P-values: 0.5363, 0.3002, 0.8416, and 0.7235 from left to right; Hunger: P-values: 0.3413, 0.8279, 0.9225, and 0.4409 from left to right). Each N represents either a group of 15 flies analyzed together in the behavioral assay (A, B, C, F) or a single fly in Dilp2 immunostaining experiments (D) and calcium imaging experiments (E). The data underlying this figure can be found in S1 Data. Data are represented as mean ± SEM with dots representing individual values and analyzed by one-way ANOVA followed by Tukey’s test (A, B, C, F) or unpaired two-tailed t test (D, E). *P (TIF)</p