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. 2017 May 2;45(11):6881–6893. doi: 10.1093/nar/gkx306

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© The Author(s) 2017. 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 4. — Analysis of non-stop decay (NSD) in ski7 and SKI complex mutants. (A) A Protein A-non-stop reporter construct was used to monitor NSD (NSD construct). This construct contains the GAL1 promoter, the Protein A coding region lacking its normal stop codon and the PGK1 3΄UTR lacking other in frame stop codons (26). A similar construct where Protein A contains its normal stop codon was used for comparison (Stop construct). (B) Protein extracts were isolated from wild-type and ski mutant strains expressing Protein A-non-stop protein and analyzed by western blotting. Strains were grown in the absence or presence of 0.5 mM H₂O₂ for 6 h. Blots were probed with a Protein A antibody (NSD) and an actin antibody as a loading control. Control denotes a wild-type strain containing an empty vector. (C) Quantification of Protein A production as shown in panel B from triplicate experiments. Error bars denote standard deviation (SD).