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. 2018 Nov 6;8:16437. doi: 10.1038/s41598-018-34773-1

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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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Microinjection of recombinant Lifeact-eGFP protein and mRNA to fluorescently label nuclei and cell outlines. (A) Zygotes injected with ~3.4 mg/ml Lifeact-eGFP protein were fixed 2, 4, 6 and 24 hpf. Maximum projections of 5 z-planes near the surface or centre of the embryo are shown. Scale = 25 μm. (B) Schematic representation of bicistronic in vitro transcribed NLS-eGFP-V2A-mCherry-CaaX mRNA: eGFP with a nuclear localization signal (NLS) is coupled to mCherry with a C-terminal CaaX box for farnesylation and insertion into the membrane. The fluorophores are separated by the self-cleaving V2A peptide. (C) mRNA-injected embryos were fixed 6 or 24 hpf or 4 days post-fertilization (dpf). Left panels show eGFP-labeled nuclei, middle panels show mCherry-labeled membranes, and the right panel shows the merged images (eGFP green; mCherry magenta). Scale = 25 μm.