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. 2012 Aug 29;9:178. doi: 10.1186/1743-422X-9-178

Figure 7.

Figure 7

3′Polyadenylation of reporter mRNA abrogates NS1-responsiveness. (A) Denaturing 1% agarose gel electrophoresis of T7 transcripts generated from XbaI digested pRL-null (Promega) and BsmBI digested pS10-RLuc. Lane 1, Rluc transcript, lane 2, Rluc transcript after 3′ polyadenylation with poly(A) polymerase, lane 3, S10-Rluc transcript, lane 4, S10-Rluc transcript after 3′ polyadenylation with poly(A) polymerase. M, 1 μg of ssRNA markers (Promega), with length indicated in nucleotides. (B) BSR 96 well monolayers transfected with 100ng pCAG NS1 and capped reporter transcripts. The amount of luciferase was assayed in triplicate 20 h after transfection with the reporter RNAs. Values are relative light units (RLU) expressed as the mean of three replicates. (C) Effect of 3′ poly(A) tail on fold upregulation of expression in response to NS1. Bars indicate one standard deviation from the mean.