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. 2019 Dec 17;9:19226. doi: 10.1038/s41598-019-55429-8

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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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Transient expression of GFP + GSF, GFP + GSF-TM and ER + RFP in tobacco cells. (A) Schematic diagram for the GFP + GSF and GFP + GSF-TM constructs transiently expressed in tobacco leaves. (B) Agrobacterium-mediated transient expression of GFP + GSF and ER + RFP in the epidermal cells of N. benthamiana. The GFP + GSF fusion protein (−3, −4) accumulated in organelle-like structures that were highly similar to the ER where the RFP was localized (−5, −6). Blue DNA-staining (Hoechst) indicated the nuclei (red arrow in −1, −2, −7 and −8). A merged fluorescence image of (−1, −3, −5) in (−7); (−2, −4, −6) in (−8) showing the similar localization of GFP + GSF and ER + RFP (white arrows in −8). (−2, −4, −6 and −8) are close-up images from (−1, −3, −5 and −7), respectively. Scale bars: 10 μm in (−1, −3, −5, −7) and 5 μm in (−2, −4, −6, −8). (C) Agrobacterium-mediated transient expression of GFP + GSF-TM and ER + RFP in the epidermal cells of N. benthamiana. The GFP + GSF-TM fusion protein (−3, −4) that accumulated in the nucleus (white arrow) was highly similar to the blue DNA-staining (Hoechst) (−1, −2) and different from where ER + RFP was localized (−5, −6). A merged fluorescence image of (−1, −3, −5) in (−7); (−2, −4, −6) in (−8) showing the similar localization of GFP + GSF-TM and blue DNA-staining (Hoechst) (white arrows in −7). (−2, −4, −6 and −8) are close-up images from (−1, −3, −5 and −7), respectively. Scale bars: 10 μm in (−1, −3, −5, −7) and 5 μm in (−2, −4, −6, −8).