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. 2018 Apr 30;8:6707. doi: 10.1038/s41598-018-25273-3

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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In vitro translation assays performed with 10 ng/assay capped or uncapped “average synthetic” RLuc, PMA1, GRE1 or SSA1 5′UTRs fused to Renilla Luciferase ORF and increasing amounts of His6x-eIF4E alone (green), eIF4E/His6x-p20 complex (red) or eIF4E/His6x-p20 6× mutant complex (blue). Data presented are the mean luminescence activity in relative light units (RLU ± s.e.m.; n = 3). The calculated free energy (kcal/mol) of each 5′UTR is shown above each panel in (a). (a) In vitro translation assays of capped RNAs. (b) In vitro translation assays of uncapped RNAs.