Figure 3.

Quantification of the level of RNAi resistance. (A) A 250 bp Nef fragment (nucleotides 8448–8698 in the LAI RNA genome) that encompasses the target and flanking regions was cloned downstream of the firefly luciferase gene in the pGL3 reporter plasmid. These reporter gene constructs were cotransfected into C33A cells with the siRNA-Nef expressing plasmid pBS-siRNA-Nef. (B) Luciferase expression observed after transfection of the reporter constructs with increasing amounts of pBS-siRNA-Nef. The level of expression observed in the absence of siRNA-Nef was set at 100% for each reporter construct. This level did not vary significantly for the different constructs. The mean values obtained in three independent experiments are shown (±SE).