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. 2018 Oct 26;10(11):585. doi: 10.3390/v10110585

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Tag orientation of a full-length fluorescent protein fusion influences the localization pattern of PLRV P3a. The panels represent single-plane confocal micrographs of N. benthamiana leaf epidermal cells co-expressing (A,B) PLRV EYFP-P3a or (C,D) P3a-ECFP with (A,C) COX4-mCherry or (B,D,E) MAN49-mCherry. EYFP/ECFP fluorescence is falsely colored green with mCherry fluorescence in red. The overlap of EYFP/ECFP and mCherry fluorescence with the corresponding brightfield image is shown in the column labeled merge with co-localization of fluorescence appearing yellow. Chloroplast autofluorescence is highlighted in blue. White arrowheads indicate EYFP fluorescence independent of COX4-mCherry. A white asterisk marks the position of the nucleus. White arrows mark MAN49-mCherry labeled cis-Golgi partially co-localizing with P3a-ECFP at putative ER exit sites. Scale bars show the length indicated.