Figure 3.

Luciferase assay. (A) Outline of luciferase assay protocol. Cells were lysed and each lysate was mixed with Dual-Luciferase Assay reagents (Promega). The first reagent initiates a reaction specific for firefly luciferase, and the second reagent both quenches the first reaction and initiates a reaction with Renilla luciferase. Light emitted by each reaction was measured for 10 s in a Turner TD20/20 luminometer. (B) Luciferase activity for selected transfected samples. From left to right, these data pairs reflect full expression of the reporter plasmid, successful knockdown of Renilla luciferase expression, the effect of transfecting a nontargeting siRNA, and the efficacy of a student-designed siRNA. Each pair represents the firefly luciferase activity in blue/green and the Renilla luciferase activity in red/orange. The ratio of Renilla to firefly luciferase activity is shown for each pair. Replicates of the indicated samples as well as no plasmid controls are not shown.