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. 2019 Sep 13;4:32. doi: 10.1038/s41392-019-0060-y

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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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Fig. 1 — E6 of HPV18 affects mRNA translation in HeLa cells. a Schematic of the protocol for identifying E6-regulated mRNA translation at the transcriptome level. b Counts of polysome-associated mRNAs (fragments per kilobase per million mapped fragments, FPKMs) from E6-silenced (shE6) and control (shControl or shCtrl) cells. mRNAs with over a two-fold decrease or increase are indicated in blue and green, respectively. c Distribution of changes in translation efficiency for shControl and shE6 cells. mRNAs with decreased (fold change ≤ −2) or increased (fold change ≥ 2) translation efficiencies are indicated as above. d, e Some mRNAs with inconsistent changes between the transcription level and translation level were further confirmed by qRT-PCR. The transcription levels (d) and translation levels (e) were measured using total RNAs or polysome-associated mRNAs. The data represent means ± SEM from at least three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001). f The protein levels of the indicated genes in shE6 and shControl HeLa cells were determined by western blot. g, h The transcription levels (g) and translation levels (h) of the indicated mRNAs with consistent changes were measured by qRT-PCR as described in d and e. The data represent means ± SEM from at least three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001). i, j Top pathways (E6-activated pathways) enriched in shControl HeLa cells from GSEA of RNA-Seq data for polysome-associated mRNAs (i) or the transcriptome (j)