Figure 7.
Triglyceride synthesis and storage in CEPT1-KO cells.A, cytosolic LDs were visualized with BODIPY 493/503 in cells treated with 300 μM oleate complexed with BSA (nuclei were stained with DAPI) (The scale bar represents 10 μm). B, synthesis of TAG was quantified after a 4 h incubation with 100 μM [3H]oleate (65 nCi/ml). Results are the mean and SD of 3 to 6 biological replicates. C, quantification of total BODIPY-positive cLDs in U2OS and CEPT1-KO cells. D, total cLD area in U2OS and CEPT1-KO cells. E, area distribution of cLDs in U2OS and CEPT1-KO cells. cLDs were binned into size groupings and expressed as a percentage of total. Results in panels B–D show the mean and SD from the analysis of 15 to 40 fields of cells (representative images are shown in panel A) from three or four biological replicates. Significance was determined by one-way ANOVA and Tukey’s multiple comparison tests (panels C and D) or unpaired t tests compared to matched U2OS controls (panels B and E). BSA, bovine serum albumin; CEPT1, choline/ethanolamine phosphotransferase 1; LD, lipid droplet; TAG, triacylglycerol.