FIG 3.

Effect of AZ-27 on RNA replication by the RSV RdRp in the minigenome system. (A) Schematic diagram of the minigenome used to measure RNA replication. In this minigenome, all transcription signals from the 3′ end, including the le and first gs signal, were removed and replaced with nucleotides 1 to 36 of the tr promoter. The trailer region at the 5′ end of the minigenome contained a deletion of the 5′ terminal 22 nucleotides to avoid terminal complementarity and to inactivate the tr promoter that typically would be present at the 3′ end of the replication product. (B and C) Effect of various concentrations of AZ-27 on minigenome template and RSV replication product in transfected cells, as determined by Northern blotting. Panel B shows the input minigenome template, and panel C shows the replication product generated by the RSV RdRp. (D) Quantification of the replication product, as determined by Northern blotting, with the numbers expressed relative to the mean levels of replicative RNA generated from the minigenome in the absence of compound. Each point represents the means from three independent experiments, with standard errors shown.