Figure 2.

BFA stimulates release of free fatty acids from stored neutral lipid droplets. (A) CHO K2 cells were incubated in the presence of 2 μg/ml BFA for the indicated time at 37°C or EtOH carrier for 12 h. Purified droplets were prepared and the lipids extracted, separated by TLC, stained with iodine, and quantified using ImageJ software. (B) CHO K2 cells were incubated in the presence of 100 μM oleate plus 12 μCi of [³H]oleate for 24 h. Cells were then washed and incubated in the presence of 2 μg/ml BFA or EtOH carrier for 12 h at 37°C. Media were collected, the cells washed and homogenized, and the radioactivity in both samples was measured. (C) The ³H lipids in the medium from the BFA-treated cells was extracted and separated by TLC along with the standards for the indicated lipids. The area in the TLC next to the indicated standard was scraped, and the amount of radioactivity was measured.