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. 2022 Sep 5;13:5228. doi: 10.1038/s41467-022-32124-3

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© The Author(s) 2022

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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Fig. 1 — Medicago truncatula roots expressing either VPY_pro:VPY-cpVenus (a–c), VPY_pro:cpVenus-VAP (d) or VPY_pro:ANK-cpVenus (e) (green) and plasma membrane (PM) and periarbuscular membrane (PAM) marker BCP1_pro:PT1-mCherry (red) colonized with Rhizophagus irregularis. Images of individual cortical cells prior to and during arbuscule development reveal the subcellular location of VPY (a) before AM fungal entry, (b) formation of fine branches, (c) fully collapsed arbuscule. Fluorescence images are projections of 10 optical sections on the z-axis taken at 0.5 µm intervals. Overlay images display both cpVenus and mCherry channels. Scale bars, 10 µm. H, hyphae; N, nucleus; P, puncta (arrow); C, crescent (arrowhead). DIC, differential interference contrast images of cells. Representative images of two independent transformations, with n = 25 arbuscule-containing cells observed in VPY-cpVenus, n = 43 arbuscule-containing cells in cpVenus-VAP, and n = 45 arbuscule-containing cells in ANK-cpVenus.