Fig. 5

NAA40 depletion in Drosophila larval fat body promotes lipogenesis and triglyceride accumulation. FB-specific Naa40 knockdown was achieved with the use of the FB-specific GAL4 promoter fly line, 0.68 Lsp2-GAL4, crossed to a UAS-Naa40RNAi line (FB > Naa40i). A–D Phalloidin staining of L3 FBs. Panels A and B outlined in yellow represent widefield images taken at × 20 and × 40 respectively of a control FB. Panels C and D outlined in red, are corresponding × 20 and × 40 images from a Naa40RNAi FB. E, F Widefield images of Nile red-stained FBs acquired at the same settings as in A–D. G, H Confocal imaging of Nile red-stained FBs. Optical stacks (3 optical sections × 0.5 μm each) of control and Naa40RNAi-stained FBs, obtained at the same settings. Scale bars A–F 200 μm and C, D 50 μm (I) FB thickness measurements of control and Naa40RNAi samples (ncontrol = 10; nFB > Naa40i = 9). J Number of lipid droplets (LDs) per fat cell in control and Naa40RNAi samples (LDs in a total of six fat cells were evaluated for three different biological samples per group). K Representative immunoblot run in triplicate of protein extracts prepared from whole L3 larvae for pAKTSer505, tAKT, β-actin in control, and Naa40RNAi conditions (n = 3/group). L RT-qPCR analysis of Lipin and Fasn in control and Naa40RNAi FBs (n = 3/group). M Triglyceride (TG) concentration (mmol/g protein) in whole L3 larvae (ncontrol = 8; nFB > Naa40i = 16). All data are presented as mean ± SEM and analysed by unpaired 2-tailed Student’s t test; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Individual values can be found in Additional file 9: Table S7