Figure 6

The LPCAT1/2 fly ortholog CG32699 resembles human LPCAT1 and 2. A) Drosophila melanogaster S2 Schneider cells were transfected with GFP-CG32699, which is the fly ortholog of mammalian LPCAT1 and LPCAT2. The growth medium was supplemented with 100 μM oleate. In the merged image, LDs are shown in red and CG32699 in green. Lower panels show a higher magnification of the central region of the pictures in the upper panels. Scalebar = 10 μm. Note the colocalization of GFP and LD540 signals. Resolution is limited due to the small size and round shape of S2 cells. B) Female virgin flies of a tubulin-Gal4 containing fly strain were crossed with male flies of three different CG32699-specific RNAi containing fly strains. L3 larvae were selected, fat bodies were isolated and representative DIC images of those fat bodies are shown for wildtype (wt), white mutant (white) and the three RNAi strains 32382, 32924 and 104570. The amount of CG32699 mRNA was measured by quantitative PCR. Scale bar = 10 μm. C + D) Bright field images as shown in panel B were quantified with ImageJ. Mean lipid droplet number per frame and mean lipid droplet size is presented for 3 frames per condition of 4 different experiments. significances were calculated by unpaired T-test analysis (*** p ≤ 0.001, ** p ≤0.01) compared to control. E) Female virgin flies of a tubulin-Gal4 containing fly strain were crossed with male flies of 3 different CG32699-specific RNAi containing fly strains or white mutant (white). From each cross three L3 larvae were selected and subjected to a LPCAT activity assay. Data were analyzed with Gel Pro Analyzer. Standard deviations and significances were calculated from three experiments by unpaired T-test analysis (** p ≤ 0.01, * p ≤0.05) compared to control (white).