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. 2019 Dec 4;10:5516. doi: 10.1038/s41467-019-13543-1

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© The Author(s) 2019

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, b Top panels: PIN2:VEN distribution in 6 DAG day-old eir1-4 PIN2p::PIN2:VEN (a) and bri1-6 eir1-4 PIN2p::PIN2:VEN (b) root meristem epidermis cells. Middle panels FM4-64 distribution after 20 min of treatment (FM4). Bottom panels: Merged images. White arrowheads: co-localization in endocytic vesicles. c PIN2:VEN signal ratio between PM and endosomal vesicles in eir1-4 PIN2p::PIN2:VEN and bri1-6 eir1-4 PIN2p::PIN2:VEN root meristem epidermis cells (n = 8 roots and 82/72 cells/dataset). Two-tailed t-test analysis of resulting values revealed a significant difference. d, e PIN2:VEN in 6 DAG eir1-4 PIN2p::PIN2:VEN root epidermis cells in mock-treated samples PNS (d) or with 100 nM eBL for 16 h (e). f Western blots performed with eir1-4 PIN2p::PIN2:VEN root membrane protein fraction grown on PNS or treated with 100 nM eBL or 28-homoBL for 16 h. Membrane protein extract from eir1-4 served as control. g, h Heat map of eir1-4 PIN2p::PIN2:mCherry root meristem epidermis cells at 6 DAG, seedlings were grown with roots in the dark, either on control medium (g) or treated with 100 nM eBL for 3 h (h). i, j Heat map of PIN2:VEN distribution in epidermis cells of 6 DAG eir1-4 PIN2p::PIN2:VEN, treated with 10 μM NAA for five hours (i) or pre-treated with 100 nM for 16 h before NAA treatment (j). k Box plot, displaying PIN2:VEN signal distribution after NAA and eBL/NAA treatments (see ‘i, j’; n = 5 roots and 39 cells/dataset). l Western blots performed with eir1-4 PIN2p::PIN2:VEN root membrane protein fraction treated with 10 μM NAA for 5 h (NAA) or pre-treated with 100 nM eBL for 16 h before NAA treatment (24-epiBL/NAA). Material from mock-treated (sol.) and eBL-treated seedlings (24-epiBL), served as controls. Anti-α-tubulin (TUB) was used as loading control. Whiskers in box plots represent the entire range of outliers; gray boxes: first and third quartiles; center line: median; dots: values obtained. Two-tailed t-test analysis has been performed to test for p-values. Scale bars: a, b = 5 μm; d, e, g–j = 10 μm. Source data are provided as Source Data file.