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. 2020 Jan 24;10:1167. doi: 10.1038/s41598-020-58082-8

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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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Self-assembling GFP analysis of non-photosynthetic Nitzschia sp. plastid TPT homologues in Phaeodactylum tricornutum. Simultaneous expression of NspTPT1 fused to GFP(S11) with a cytosolic GFP(S1–10) led to a cER-characteristic fluorescence pattern (A). No fluorescence could be observed after expression of NspTPT1:GFP(S11) with the ER/cER marker (PDI) fused to GFP(S1–10) (B). Fluorescence signals were obtained upon expression of the NspTPT2:GFP(S11) fusion proteins with both the ER/cER- (C) and the PPC-marker fused to GFP(S1–10) (D). Both NspTPT4a:GFP(S11) (E) and NspTPT4b:GFP(S11) (G) did not show a fluorescence signal when expressed with the IMS marker MGD1 fused to GFP(S1–10), whereas a clear GFP signal circling the plastid autofluorescence could be observed when NspTPT4a:GFP(S11) (F) and NspTPT4b:GFP(S11) (H) were expressed with the stromal-targeted AtpC:GFP(S1–10). TL, transmitted light; PAF, plastid autofluorescence; GFP, enhanced green fluorescent protein; PAF/GFP, overlay of plastid and GFP fluorescence; Cyt., cytosol; cER, chloroplast endoplasmic reticulum; PPC, periplastidal compartment; IMS, intermembrane space; Str., plastid stroma; scale bar represents 10 µm.