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. 2020 May 19;10:8286. doi: 10.1038/s41598-020-64623-y

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© The Author(s) 2020

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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MACROD1, MACROD2 and TARG1 are differentially localised. (a) HeLa Flp-In T-REx cells overexpressing MACROD1, MACROD2 or TARG1 were treated overnight with 100 ng doxycycline/ml, fixed with PFA, stained with primary antibodies (monoclonal antibodies used: MACROD1 (28C11), MACROD2 (18D12), TARG1 (3A5)), visualised using an AlexaFluor488-coupled secondary antibody and analysed with confocal microscopy. (b) HeLa Flp-In T-REx control cells were treated and analysed as in panel (a). (c) HeLa cells were transfected with the indicated N- or C-terminally GFP-tagged constructs and analysed using live-cell confocal microscopy. (d) HeLa cells were transfected with constructs expressing mTurquoise2 targeted to mitochondria and either mRuby2-labeled MACROD1 full-length construct or an N-terminal truncation lacking amino acids 1–77, followed by live-cell confocal imaging. Scale bars represent 10 µM.