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. 2020 Aug 27;11:4285. doi: 10.1038/s41467-020-17949-0

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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 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. 4 — a–f Co-localization of CRE1/AHK4-GFP with the plasma membrane markers PIP1;4-mCherry (a, b), NPSN12-mCherry (c, d) and FM4-64 (e, f) in the root meristematic epidermal cells. Profiles of fluorescence intensity of PM marker (red line) and CRE1/AHK4-GFP (green line) (b, d, f) were measured along the white lines (a, c, e) starting from the upper end (0 µm) towards the arrowhead. Peaks of PIP1;4-mCherry and NPSN12-mCherry fluorescence maxima correlate with the plasma membrane staining and overlap with CRE1/AHK4-GFP signal maxima (black arrows) (a–d). CRE1/AHK4-GFP signal detected at the cell plate of dividing cell (black stars on c, e) co-localizes with FM4-64 marker (e, f; black stars) but not with the plasma membrane marker NPSN12-mCherry (c, d; black stars). g Super-resolution imaging (SIM) of CRE1/AHK4-GFP subcellular co-localization with FM4-64 labelled PM. h, i Co-localization of CRE1/AHK4-GFP and FM4-64 (h), but not p24δ5-RFP (i) in endosomal compartments (blue arrowheads) formed in root meristematic epidermal cells treated with 50 µM in BFA for 1 h. Note CRE1/AHK4-GFP localization at the plasma membrane (white arrows) prior BFA treatment. Scale bars = 5 μm (a, c, e, h, i) and 2 μm (g).