PC-specific downregulation of upd3 expression and JNK signaling result in a rescue in the elevated levels of prc expression in the fat cells of HSD-fed flies. (A) Expression of dot-Gal4 driving UAS-GFP (green) specifically in the PCs adjacent to the cardiac tube (red; stained with phalloidin). The abdominal body wall muscles and the alary muscles are also stained with phalloidin red. (B) Increase in the levels of prc expression in fat cells of the flies of the genotypes mentioned when reared on HSD. The transcript levels are normalized to that of the constitutive ribosomal gene rp49. (C–E) Changes in heart period (C), diastolic interval (D), and systolic interval (E) upon knocking down upd3 in the PCs of HSD-fed flies. The dots represent the samples analyzed for each genotype. (F and G) Drop in the reporter GFP expression for gstD1 in the PCs of HSD-fed flies (G) as compared to those reared on ND (F). Phalloidin (red) marks the cardiac tube and the alary muscles. (H) Quantification of the mean fluorescence intensity for gstD1-GFP expression in the PCs of flies reared on either ND or HSD. The dots represent the number of PCs analyzed for each genotype. (I) Feeding on HSD leads to an increase in the levels of upd3 expression in the PCs of flies of all the genotypes mentioned. The transcript levels are normalized to that of the constitutive ribosomal gene rp49. (J) Increase in the levels of upd3 expression in the PCs of flies of the genotypes mentioned when reared on HSD. The transcript levels are normalized to that of the constitutive ribosomal gene rp49. Genotypes are as mentioned. Data are represented as mean ± SD. P values (ns ≥ 0.05, **P < 0.01, ***P < 1 × 10−3) were obtained by unpaired Student’s t test (two-tailed) with Welch’s correction (H), by one-way ANOVA with Tukey’s multiple-comparison test (C, D, and E), and by two-way ANOVA with Tukey’s multiple-comparison test (B, I, and J). Scale bars, 100 μm (A) and 50 μm (F and G).
