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. 2016 Sep 7;45(3):1416–1432. doi: 10.1093/nar/gkw795

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 9. — Functional analysis of the 3΄UTR sequences predicted to base pair with the IRES element. (A) Diagram of the base pairs disrupted between the domain 4 or domain 5 of the IRES and the 3΄UTR in constructs SL1a or Ha. Red letters depict nucleotide substitutions of the 3΄UTR sequence. (B) Translation efficiency of the SL1a and Ha constructs. The Firefly luciferase (Fluc) activity/μg of protein and Renilla luciferase (Rluc) activity/μg of protein was made relative to the values obtained for the wild type construct performed in parallel. Relative IRES activity (mean ± SD) was determined in transfected BHK-21 cells as the Fluc/Rluc ratio expressed from monocistronic constructs, then normalized to the activity observed for the wild type IRES (set to 1). Each experiment was performed in triplicate and repeated three times.