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. 2018 Jan 2;9:10. doi: 10.1038/s41467-017-02449-5

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© The Author(s) 2017

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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Fig. 3 — The Poly(A) SPOT-ON attenuates nascent protein synthesis. a SUnSET measurements in U2OS cells were conducted in the absence of puromycin to determine background levels of signal. Puromycin staining (green), phallodin (red), DNA (blue), and the merge between channels are arranged from top to bottom. As a positive control, puromycin and vehicle were used to determine the upper limit of translation. Both homoharringtonine (HHT) and the Poly(A) SPOT-ON robustly decrease protein synthesis, whereas the scrambled SPOT-ON failed to do so. b Quantification of a, empty boxes indicate no puromycin control, pink boxes are the positive control, blue boxed are homoharringtonine, green boxes are the Poly(A) SPOT-ON, and purple boxes are the scramble control. n = 15. c PABP overexpression rescues decreased protein synthesis caused by the Poly(A) SPOT-ON. Drug treatments consisted of either vehicle or SPOT-ON in the presence of an empty vector or overexpressed PABP. The amount of vector is indicated above the row of images. Markers are arranged as in a. d Quantification of c. n = 6. Columns represent measurements in the same manner as in b. *P < 0.05, **P < 0.01, significantly different from vehicle+puro group analyzed by one-way ANOVA followed by Bonferroni post hoc test. For all graphs shown in the figure, data are plotted as mean ± s.d