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. 2019 Dec 24;9:19812. doi: 10.1038/s41598-019-56231-2

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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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Northern blot of subcellular localization of mitosRNA-tRNA-cys 3′ variants in the WS40NE cell line. 5 µg RNA was loaded in each well and blots were hybridized with 0.5 nM mitosRNA-tRNA-cys probe. Cells were treated with anoxia for 24 or 49 h. (a) mitosRNAs derived from tRNA-cys, around 20–30 nucleotides long, are enriched in the mitochondrial fraction and in response to anoxia compared to whole cells. Note, mitosRNA band is not very strong in the northern blot of the whole cells here (4a) while they are clearly present in 3b. This discrepancy is due to the difference in loading 5 µg RNA for Fig. 5a vs 20 µg RNA for Fig. 4b. (b) Subcellular fractionation suggests the mitosRNAs are distributed in the mitochondrial and cytoplasmic compartments. N = normoxia; A = anoxia; Cytoplasm = non-mitochondrial cytoplasmic fraction.