Figure 4

Mdm2 inhibits NF-Y mediated hTR transactivation. 5637 cells were co-transfected with 1.5 μg of the pLh2023(-796/+69) construct, 0.5 μg of expression vectors encoding wild-type NF-YA, B and C together with increasing amounts of mdm2 expression vector (from 0 to 2.5 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 the cell lysate were assayed for firefly and Renilla luciferase activity. The data is expressed as fold induction of luciferase activity relative to the promoter alone and the means of the three independent experiments performed in duplicate.