Figure 6.
Increased levels of upd3 expression in the PCs of HSD-fed flies is dependent on lipid uptake and lipolysis. (A–D) Changes in the levels of neutral lipid accumulation (green; BODIPY staining) in the PCs of HSD-fed flies (B) and upon downregulation of either LpR1 (C) or LpR2 (D) in the PCs of HSD-fed flies as compared to that observed in ND-fed flies (A). (E) Quantification of the mean fluorescence intensity for BODIPY staining in the PCs of flies reared on ND, HSD, and upon downregulation of LpR1 or LpR2 in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (F–I′) Changes in the levels of Nile Blue A staining in the PCs of HSD-fed flies (G and G′) and upon downregulation of either LpR1 (H and H′) or LpR2 (I and I′) in the PCs of HSD-fed flies as compared to that observed in ND-fed flies (F and F′). F, G, H, and I Show Nile Blue A expression in grayscale; F′, G′, H′, and I′ represent the same images in blue channel. (J) Quantification of the mean fluorescence intensity for Nile Blue A staining in the PCs of flies reared on ND, HSD, and upon downregulation of either LpR1 or LpR2 in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (K) Changes in the levels of upd3 expression in the PCs of flies reared on HSD, and upon downregulating either LpR1 or LpR2 in the PCs of HSD-fed flies as compared to that observed in flies reared on ND. The transcript levels are normalized to that of the constitutive ribosomal gene rp49. (L–N) Decrease in the levels of neutral lipid accumulation (BODIPY staining; green) in the PCs of HSD-fed flies upon overexpression of bmm (N) as compared to that observed in HSD-fed flies (M) and in ND-fed flies (L). (O) Quantification of the mean fluorescence intensity for BODIPY staining in the PCs of flies reared on ND, HSD, and upon overexpression of bmm in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (P–Q′) Increase in the levels of Nile Blue A staining in the PCs of HSD-fed flies upon overexpression of bmm (Q and Q′) as compared to that observed in HSD-fed flies (P and P′). P and Q show Nile Blue A expression in grayscale; P′ and Q′ represent the same images in blue channel. (R) Quantification of the mean fluorescence intensity for Nile Blue A staining in the PCs of flies reared on HSD, and upon overexpression of bmm in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (S) Increase in the levels of upd3 expression in the PCs of flies reared on HSD, and upon overexpression of bmm in the PCs of HSD-fed flies as compared to that observed in flies reared on ND. 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 (R) or by two-way ANOVA with Tukey’s multiple-comparison test (E, J, K, O, and S). Scale bars, 50 μm.

Increased levels of upd3 expression in the PCs of HSD-fed flies is dependent on lipid uptake and lipolysis. (A–D) Changes in the levels of neutral lipid accumulation (green; BODIPY staining) in the PCs of HSD-fed flies (B) and upon downregulation of either LpR1 (C) or LpR2 (D) in the PCs of HSD-fed flies as compared to that observed in ND-fed flies (A). (E) Quantification of the mean fluorescence intensity for BODIPY staining in the PCs of flies reared on ND, HSD, and upon downregulation of LpR1 or LpR2 in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (F–I′) Changes in the levels of Nile Blue A staining in the PCs of HSD-fed flies (G and G′) and upon downregulation of either LpR1 (H and H′) or LpR2 (I and I′) in the PCs of HSD-fed flies as compared to that observed in ND-fed flies (F and F′). F, G, H, and I Show Nile Blue A expression in grayscale; F′, G′, H′, and I′ represent the same images in blue channel. (J) Quantification of the mean fluorescence intensity for Nile Blue A staining in the PCs of flies reared on ND, HSD, and upon downregulation of either LpR1 or LpR2 in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (K) Changes in the levels of upd3 expression in the PCs of flies reared on HSD, and upon downregulating either LpR1 or LpR2 in the PCs of HSD-fed flies as compared to that observed in flies reared on ND. The transcript levels are normalized to that of the constitutive ribosomal gene rp49. (L–N) Decrease in the levels of neutral lipid accumulation (BODIPY staining; green) in the PCs of HSD-fed flies upon overexpression of bmm (N) as compared to that observed in HSD-fed flies (M) and in ND-fed flies (L). (O) Quantification of the mean fluorescence intensity for BODIPY staining in the PCs of flies reared on ND, HSD, and upon overexpression of bmm in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (P–Q′) Increase in the levels of Nile Blue A staining in the PCs of HSD-fed flies upon overexpression of bmm (Q and Q′) as compared to that observed in HSD-fed flies (P and P′). P and Q show Nile Blue A expression in grayscale; P′ and Q′ represent the same images in blue channel. (R) Quantification of the mean fluorescence intensity for Nile Blue A staining in the PCs of flies reared on HSD, and upon overexpression of bmm in the PCs of HSD-fed flies. The dots represent the number of PCs analyzed for each genotype. (S) Increase in the levels of upd3 expression in the PCs of flies reared on HSD, and upon overexpression of bmm in the PCs of HSD-fed flies as compared to that observed in flies reared on ND. 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 (R) or by two-way ANOVA with Tukey’s multiple-comparison test (E, J, K, O, and S). Scale bars, 50 μm.

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