FIG. 2.
Quantification of expression from reporter plasmids does not require normalization with a Renilla luciferase construct. (A) A total of 100 ng pGL3 and 1.5 μg pBluescript carrier was transfected into each well of a six-well plate. Luciferase expression from each well was measured and normalized to the expression of the first well. Variation was tested using a single sample t test (H0 = 1) to determine whether any given well varied statistically from the first. No statistically significant variation was observed (P ≫ 0.05). (B) Cells were transfected with 100 ng of biotinylated (white bars) or unbiotinylated (black bars) pGL3-p21waf (reporter), 50 ng SV40 promoter-driven Renilla luciferase (normalization protein), and increasing amounts of the BBP-PML IV plasmid (targeting protein). The top and bottom graphs were derived from the same data, whereas the data for the bottom graph were normalized to that of Renilla luciferase. Error bars in panels A and B represent the standard deviations of three separate measurements made from each well.