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. 2022 Oct 6;11:e80282. doi: 10.7554/eLife.80282

Figure 6. Manipulating Pect levels in the fat body alters phospholipid profile.

(A) Average concentrations of phosphatidylcholine (PC), phosphatidylethanolamine (PE), lysophosphatidylcholine (LPC), and lysophosphatidylethanolamine (LPE) lipid classes in LppGal4>UAS-Pect-RNAi flies compared to control under 7-day normal food (NF) conditions. (B) Depicts the concentration of Pect-associated phospoholipids in LppGal4>UAS-Pect-RNAi flies compared to control. Lipidomics was performed using a targeted quantitative lipidyzer (Sciex 5500 Lipidyzer). 4–6 independent biological replicates were used for each genotype, with n = 10 flies composing one biological replicate. Unpaired t-test with Welch’s correction. Asterisks indicate significant changes with p-value<0.05, p-value<0.005, and p-value<0.0005. Error bars = standard deviation, points = individual replicate values.

Figure 6—source data 1. Lipidomics for fat-specific Pect knockdown and Pect overexpression on normal food.
Relevant to Figure 6, Figure 6—figure supplements 2–4.
elife-80282-fig6-data1.xlsx (400.5KB, xlsx)

Figure 6.

Figure 6—figure supplement 1. Pect mRNA expression.

Figure 6—figure supplement 1.

(A) Mean fold change in Pect mRNA levels in the fat-specific (left) Pect knockdown flies and (right) Pect overexpression flies. Unpaired t-test with Welch’s correction. (B) Mean fold change in Pect mRNA levels of normal food (NF)-fed and high-sugar diet (HSD)-fed flies overtime. Two-way ANOVA with Holm–Sidak correction. N = 3 technical replicates/group of cDNA collected from an N = 30 flies/group. Asterisks indicate significant changes with p-value<0.05, p-value<0.005, and p-value<0.0005. Error bars = standard deviation.
Figure 6—figure supplement 1—source data 1. Pect expression in RNAi and OE lines related to Figure 6—figure supplement 1A.
Figure 6—figure supplement 1—source data 2. Pect expression in RNAi and OE lines related to Figure 6—figure supplement 1B.
Pect expression in w1118 at different normal food (NF) and high-sugar diet (HSD) timepoints related to Figure 6—figure supplement 1B.
Figure 6—figure supplement 2. Fourteen days of high-sugar diet (HSD) cause an increase in Pect-associated phospholipid classes.

Figure 6—figure supplement 2.

Percent lipid composition for all (A) phosphatidylcholine (PC) and (B) phosphatidylethanolamine (PE) classes in normal food (NF)-fed flies overtime. Note that the overall composition does not change with age. The lipid composition was averaged amongst 10 biological replicates (n = 10 flies/replicate). Error bars indicate standard deviation. (C) Average concentration of PC and PE classes that are associated with Pect based on van Dam et al., 2020. Bars plot average concentration amongst 10 biological replicates (n = 10 flies/replicate). Two-way ANOVA with Holm–Sidak correction. Asterisks indicate significant changes with p-value<0.05, p-value<0.005, and p-value<0.0005. Error bars indicate standard deviation.
Figure 6—figure supplement 3. Additional lipid class responses to fat body Pect knockdown.

Figure 6—figure supplement 3.

Average concentrations of triacylglyceride (TAG), CE, diacyl glyceride (DAG), SM, and free fatty acid (FFA) lipid classes in LppGal4>UAS-Pect-RNAi flies compared to control under 7-day normal food (NF) conditions. Lipidomics was performed using a targeted quantitative lipidyzer (Sciex 5500 Lipidyzer). 4–6 independent biological replicates were used for each genotype, with n = 10 flies composing one biological replicate. Unpaired t-test with Welch’s correction. Asterisks indicate significant changes with p-value<0.05, p-value<0.005, and p-value<0.0005. Error bars = standard deviation.
Figure 6—figure supplement 4. Lipidomic profile of fat body Pect overexpression flies.

Figure 6—figure supplement 4.

Average concentrations of phosphatidylcholine (PC), phosphatidylethanolamine (PE), CE, lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), SM, triacylglyceride (TAG), diacylglyceride (DAG), and free fatty acid (FFA) lipid classes in LppGal4>UAS-Pect flies compared to control under 7-day normal food (NF) conditions. Lipidomics was performed using a targeted quantitative lipidyzer (Sciex 5500 Lipidyzer). 4–6 independent biological replicates were used for each genotype, with n = 10 flies composing one biological replicate. Unpaired t-test with Welch’s correction. Asterisks indicate significant changes with p-value<0.05, p-value<0.005, and p-value<0.0005. Error bars = standard deviation.