Luciferase reporter assay using minimal promoter sequence of K-RTA and K8 binding sequence.

Relative fold increase of: (A) pGL4.23-RB, and (B) pGL4.23-RV luciferase activity in the presence of increasing amounts of K-RTA. Three copies of K-RTA binding motif RB or RV sequence was placed upstream of a minimal TATA promoter (minP) in a luciferase vector and transiently expressed in HEK293 cells along with the K-RTA protein. Relative luciferase units were calculated relative to the basal promoter activity. The lower panels are western blots showing relative expression of K-RTA and GAPDH from equal amounts of cell lysates from each experimental set. Relative fold increase of: (C) pGL4.23-KB luciferase activity in presence of increasing concentration of K8. Three copies of K8 binding motif KB sequence was placed upstream of a minimal TATA promoter (minP) in a luciferase vector pGL4.23-KB and transiently expressed in HEK293 cells along with K8. Relative luciferase units were calculated relative to the basal promoter activity. The lower panels are western blots showing relative expression of K8 and GAPDH from equal amounts of cell lysates from each experimental set.