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. 2018 Jul 2;9:2558. doi: 10.1038/s41467-018-05007-9

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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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Fig. 2 — Dysfunction of the degradosome or GRSF1 upregulates RNAs originating from L-strand transcription. a–d Tracks of mapped reads of long transcript libraries. RNA from two set of experiments (a–b and c–d) performed in triplicate was analyzed. Panels b and d show the D-loop region with an adjusted scale. siRNA silencing of endogenous PNPase expression was rescued by expression of an exogenous siRNA-insensitive version of PNPase-FLAG. tRNA genes are labeled with single-letter symbols. S1 and S2 correspond to tRNA-Ser(UCN) and tRNA-Ser(AGY), respectively. e Western blot analysis of indicated proteins. Exogenous, siRNA-insensitive PNPase was detected with anti-FLAG antibodies. Membranes were stained with Ponceau S as a loading control. f, g Contribution of L-strand templated transcripts to the mitochondrial transcriptome. Individual values are shown. Horizontal lines represent mean values. h–i Distribution of GRSF1-binding sites across mitochondrial transcripts. eCLIP data from ENCODE database were re-analyzed. The D-loop region is shown with an adjusted scale (i)