Metabolic modification of cell surface Fas expression. Changes in expression of Fas caused by removing glucose or adding insulin to the culture medium. The glucose is removed by incubating HL60 (human promyelocytic leukemia) cells in glucose free RPMI with the addition of 5 mM 2-Deoxyglucose. When cells were incubated with insulin, normal RPMI medium conditions were used with the addition of 100 μg/mL of insulin. The above conditions represent a 24 hour treatment period. This data is representative of five separate experiments. Each of the experiments showed the same general trends, however the experiments were done at different times and because the intensity of the fluorochromes varies with time, this makes direct statistical comparisons suspect. The level of Fas was detected using PE-conjugated anti-humanFas (CD95) antibodies (Pharmingen, California) and measured using a Coulter Elite Epics Flow Cytometer (Coulter, Hialeah, Florida) and FlowJo analysis software (Tree Star, Inc, Oregon).