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. 2019 Feb 4;9:1265. doi: 10.1038/s41598-018-37666-5

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© The Author(s) 2019

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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CMLD011580 targets eIF4A1. (a) Dose-dependent inhibition of protein synthesis in vivo by CMLD011580 is eIF4A1-dependent. Cells were incubated with the indicated concentrations of compound for 1 hour, and [³⁵S]-methionine was added 15 minutes before the end of the incubation. The rate of [³⁵S]-methionine incorporation is expressed relative to that of cells treated with vehicle (DMSO). Results are expressed as the mean ± SEM of 3 biological replicates. (b) Cellular toxicity of CMLD011580 is eIF4A1-dependent. Cells were incubated in the presence of the indicated concentrations of CMLD011580 or (−)-CR-1-31-b for 48 hrs, after which the percentage of viable cells was measured. Results are expressed as the mean ± SEM of 3 biological replicates. (c) Stimulation of eIF4A1:RNA binding by (−)-CR-1-31-b or CMLD011580. Fluorescently labelled RNA (16-mers) containing (AG)₈ or (UC)₈ repeats were used as probes in these experiments. Shown is the change in fluorescent polarization obtained in the presence of indicated compound concentration, above what was observed in the presence of DMSO. Results are expressed as the mean ± SEM of 3 biological replicates.