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. 2014 Jul 10;42(14):8928–8938. doi: 10.1093/nar/gku608

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© The Author(s) 2014. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — (A) Sequence boundaries of seven readthrough candidates cloned into a dual-luciferase plasmid. Main ORF stop codons are in red font and the conserved CTAG motif is highlighted in bold and underlined. Plasmids were transfected into HEK-293T cells and 24 hr later lysates analysed for readthrough by both western blotting with anti-Renilla (B) and dual-luciferase assay to determine readthrough efficiencies (C). For (B) each TGG in-frame control (I) is adjacent to its respective test (T) construct. For (C) readthrough efficiencies were calculated as the ratio of test construct luciferase activity to its TGG in-frame control from three independent experiments.