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. 2009 Mar 18;83(11):5846–5853. doi: 10.1128/JVI.02602-08

FIG. 2.

FIG. 2.

The ICR from the MDV-1 bicistronic transcript has IRES activity in a dual-luciferase reporter assay. (A) Bicistronic luciferase constructs used for transfection. The sequence (ICR) to be tested for IRES activity was inserted between the Renilla (R-Luc) and firefly (F-Luc) luciferase genes in the MCS spacer. In the psiRF-ICR/pLess construct, the SV40 promoter was removed. (B) Results of luciferase assay using DNA transfection of DF-1 and HEK 293T cells. The F:R ratio for each DNA construct was normalized to that obtained with the psiRF vector containing the MCS spacer as a negative control, whose F:R was set to 1. (C) Same as panel B, but the transfection was done with RNAs produced by in vitro transcription. (D) shRNA knockdown of the ICR. The data are results of cotransfection experiments with DF-1 cells. The firefly and Renilla luciferase activities were determined and expressed as percentages of the activity with control nonsilencing shRNA (pChU6/NS-shRNA). pChU6/sh refers to the plasmid encoding the siRNA that targets the ICR from nucleotides 131150 to 131170 (GenBank accession number AF243438).