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. 2023 Nov 30;21(4):648–656. doi: 10.1038/s41592-023-02085-6

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© The Author(s) 2023

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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Extended Data Fig. 7 — HeLa cells stably expressing td5StayGold(c4)=GianCreg were chemically fixed and immunostained for three Golgi markers: GM130 (cis), Giantin (medial), and TGN46 (trans-Golgi network). Their immunosignals (Alexa Fluor 647) were imaged with td5StayGold fluorescence three-dimensionally with a z-step of 0.52 µm using a confocal microscope. For a scan format of 512 × 512 and a zoom factor of 4×, the pixel size was 0.104 µm, which was sufficiently small to make good use of the resolving power of the high–numerical-aperture (NA 1.2) objective lens. a–c, Analysis of colocalization between td5StayGold(c4)=GianCreg and GM130 (a), Giantin (b), or TGN46 (c) in a single experiment. left, Representative single-slice images. Nuclei were stained with DAPI. Scale bar, 5 µm. right, Pearson correlation plots based on 26 (a and b) and 27 (c) z-slices, showing the correlation coefficients: 0.324 (a), 0.606 (b), and 0.325 (c). d, The experiment was repeated three times. Individual correlation coefficients are plotted; the data plots derived from a–c are highlighted in magenta). The mean values are shown by gray bars.

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