Fig. 9

Preventing excessive fat body lipid catabolism rescues the behavioral effect triggered by osi downregulation. A–D Quantitation of Lip3 and pepck mRNA levels in mutants (A, B) or larvae with ubiquitous RNAi expression (C, D). Data shows a clear increase in Lip3 with no effect on pepck levels. The total number of replicas/genotype is included; each dataset consisted of 20 larvae pooled together. A Shapiro-Wilk test was used for normality assessment; if normality was confirmed, a one-way ANOVA with Bonferroni’s multiple comparisons test was performed. For nonparametric assessment, Kruskal-Wallis with Dunn’s multiple comparisons test were performed (see Additional file 2). E qPCR analysis on RNA extracted from whole larvae shows efficient reduction of Lip3 levels when Lip3^RNAi is expressed under pplG4 at 96h AEL. The total number of replicas/genotype is included; each dataset consisted of 20 larvae pooled together. A two-tailed t-test was performed. F Lip3 downregulation partially rescues osi^RNAi lethality. Six independent experiments were performed; the total number of animals assessed are indicated. G–I Confocal images showing fat body cells stained with BODIPY (golden lipid droplets) for control, osi^RNAi, Lip3^RNAi, and Lip3^RNAi, osi^RNAi expressing animals under pplG4 from 72 and 96h AEL. Bars represent 20 μm (I). The percentage of area covered by lipid droplets or “reactive area” is quantitated in G for 72h and H for 96h AEL. Each dot represents the mean area of each fat body analyzed. The total number of fat bodies per condition is indicated. All panels: Shapiro-Wilk test was used for normality assessment. Graphs display mean ± SEM. Different letters indicate significant differences, p<0.05; treatments sharing any letter are not statistically different. The total number of samples analyzed is described below the corresponding panel. All datasets and statistical analysis on which the conclusions are drawn are included in Additional file 2