MethodsThe experiment was designed in order to have a control and one tested condition. The condition that was tested was a Linolenic acid solution. The negative control was simply a 50% ethanol solution. The acid solution was prepared in 50% ehtanol then added to a 6:1 molar ratio of acid-BSA. About 10,000 HeLa cells were plated and grown for 24 hours before being cultured in the wells. 400 micromolar of Linolenic acid or 50 percent ethanol were exposed to the cells. The cells were then incubated for 24 hours. Next, the cells were stained following manufacturer’s instructions for Lipid Spot 488, [Biotium #70065], [CF594WGA], [Biotium # 29023-1], and [Invitrogen #R37605]. All cells were washed using HBSS medium. The number of lipid droplets per cell was then counted using Image J analysis. Statistical values were determined; averages, standard deviation, standard error, and student’s t-test were calculated. |