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. 2020 Sep 2;10:14437. doi: 10.1038/s41598-020-71474-0

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Spatiotemporal characteristics of dGRAPHIC. (A) Confocal images reveal subcellular distribution of reconstituted GFP in co-cultured NT-probe-expressing and CT-probe-expressing LLCPK1 cells. Schema of z-positions of confocal images. (B) Stacked confocal images (z = 1.2 μm) at apical surface (top), middle position (center), and basal membrane (bottom). (C) Full stacked image (z = 7.2 μm) of reconstituted GFP, nuclei of NT-cells (red), and nuclei of CT-cells (blue). Scale bar, 20 µm. (D) Time lapse images for the generation of reconstituted GFP signal between NT-LLCPK1 cells (red nuclei) and CT-LLCPK1 cells (blue nuclei). Upper panels are bright field (differential interference contrast) images of the bottom fluorescence images. The reconstituted GFP signal was generated at cell–cell contact sites and gradually spread out over the whole plasma membrane of contacted NT cells (red nuclei). Scale bar, 100 μm.