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. 2018 Jul 6;8:10279. doi: 10.1038/s41598-018-28188-1

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© The Author(s) 2018

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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PIN3 phosphorylation is required for PIN3 polarization and root gravitropic response. (a) Root bending kinetics of PIN3 mutant variants during gravitropic response. Root curvatures were measured every 4 hours. Student’s T-test was calculated for the comparison of each line with the control (Col-0). PIN3-YFP-P2D mutant variant shows slower root bending. (b) Quantification of PIN3 polarization in columella cells before and after gravistimulation. Signal before gravistimulation was normalized to 1. Student’s T-test was calculated for the comparison of each line with the control (PIN3::PIN3-YFP). Both phosphomimic variants, PIN3-P1D and PIN3-P2D, exhibit less gravity-induced PIN3 relocation. (c–l) Localization of PIN3-YFP (c,d), PIN3-YFP-P1A (e,f), PIN3-YFP-P1D (g,h), PIN3-YFP-P2A (i,j), and PIN3-YFP-P2D (k,l) before (upper row) and after 30 minutes (lower row) of gravitropic stimulation in the wild type background. Values are the average of three biological replicates (n > 10 per time point on each replicate). Error bars represent SE, (*p < 0.05, **p < 0.01, ***p < 0.001). Yellow arrows indicate gravity vector. Bars = 10 µm.