Figure 2

mdm2 represses hTR promoter activity. The pRB/p53 negative cell line 5637 were co-transfected with 1.5 μg of the pLh2023(-796/+69) construct and increasing amounts of mdm2 expression vector (from 0.5 to 4.0 μg). Total input DNA amount for transfection was adjusted with Salmon Sperm DNA to ensure a constant amount in all transfections. After 48 hour of culture, cells were harvested, and cell lysates were assayed for firefly and Renilla luciferase activity. The data is expressed as fold induction of luciferase activity relative to the promoter alone and data presented are the mean of three independent experiments performed in duplicate.