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. 2018 Nov 28;8:17435. doi: 10.1038/s41598-018-35753-1

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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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Stimulation of translation by PABP in the reconstituted cap-dependent translation system. (A) Rluc, Rluc-A, Cap-Rluc or Cap-Rluc-A RNA (0.1 μM each) was translated in the reconstitution system with or without PABP (1.92 μM). After translation, samples were analyzed by western blotting with an anti-myc antibody (left panel) and Rluc assay (right panel; each column and bar represent the mean and standard deviation of three experiments, respectively). eIF4G (84–1599), which contains the PABP binding site²⁴, was used as eIF4G. (B) Cap-Rluc or Cap-Rluc-A (0.1 μM each) RNA was translated in the reconstitution system with or without PABP or a PABP mutant (M161A)⁹ (1.92 μM, each) (left panel). These RNAs were also translated with eIF4G (84–1599) or eIF4G (197–1599)²⁴ in the presence or absence of PABP (1.92 μM) (right panel). After translation, Rluc activity was measured. Each column and bar represent the mean and standard deviation three experiments, respectively; the average Rluc activity without PABP was set at 1.0.