Cholesterol content of isolated lipid droplets was determined as described
in [32]. In brief, lipids were extracted as described above and
loaded on a TLC plate alongside known amounts of cholesterol and cholesterol
linoleate standards. The plate was developed in a single phase
hexane:ether:formic acid (80:20:2). Lipids were visualized by spraying
the plate with 0.05% ferric chloride, 5% acetic and 5% sulfuric acids and
heating at 160 °C for 5–10 min, when the presence of cholesterol and
cholesterol esters is indicated by the appearance of red-violet colored
band. A standard curve of pixel intensity was generated for the cholesterol
standards using Photoshop and used to quantify the amount of
cholesterol in the lipid droplet samples. To determine the levels of