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. 2017 Nov 20;7:15833. doi: 10.1038/s41598-017-16177-9

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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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Polysome associated PTC-containing mRNAs expressed from gBglo-P66 are induced by UPF1 depletion. (a,g) Absorbance at 254 nm (upper panels) and semi-qPCR analysis of relative mRNA levels (lower panels) of sucrose gradient fractions containing polysomes of HeLa cells transfected with the indicated constructs in the absence (a–e) or presence (f and g) of siRNA targeting UPF1. Cells were treated with puromycin before they were harvested (c–g). (h) Comparison of distribution patterns of PTC-containing mRNAs in cells transfected with the indicated constructs in the presence and absence of UPF1 knockdown and shown in panels d–g. mRNA bands obtained from semi-qPCR were quantified by densitometry. Every experiment in Fig. 4 was performed two independent times and the results of one representative experiment are shown. Error bars represent the SD of the mean of semi-qPCR results using samples obtained from the two independently performed polysome analyses.