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. 2018 Feb 28;9:875. doi: 10.1038/s41467-018-03256-2

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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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Fig. 8 — : ARF5 binds to the WOX4 promoter. a–c Confocal analysis of stem bases of plants carrying the pWOX4:YFP and the pARF5:mCherry reporter. Asterisks mark the vascular bundles. Size bars represent 100 µm. d, e Analysis of WOX4 transcript levels by quantitative RT-PCR at the stem base of wild-type and mp-S319 mutant pants (d) and in the second internode of pPXY:GR-ARF5∆III/IV plants after mock or Dex and CHX or CHX/Dex (e) treatments. Student’s T-test (wild type and mp-S319 p = 1.74E-02 and Mock and Dex p = 1.66E-02) or Welch’s T-test (CHX and CHX/Dex p = 1.29E-02) were performed comparing wild-type and mp-S319 mutants, mock and Dex and CHX and CHX/Dex, respectively (Sample sizes n = 3–6). Error bars represent ±standard deviation. Significance is indicated by the asterisk. f Analysis of relative luciferase activity of a pWOX4:LUC (firefly);p35S:LUC (Renilla) reporter in Arabidopsis protoplasts in the presence of p35S:ARF3, p35S:ARF5∆III/IV or no effector construct. Relative luciferase activity was determined by dual-luciferase assays. Statistical groups indicated by letters were determined by one-way ANOVA with post hoc Tamhane-T2 (CI 95%, Sample size n = 4–5). The T-bars that extend from the boxes (whiskers) are expected to include 95% of the data. g EMSAs probing DNA oligomers with proven (PIN1, STOMAGEN)^50,53 or predicted (WOX4) binding capacities by the ARF5 DNA-binding domain (ARF5-DBD). An oligomer from the CLAVATA1 (CLV1) promoter was used as a negative control. Oligomer sequences are shown in Supplementary Table 1. h EMSAs probing the WOX4a oligomer in the presence of the non-labelled WOX4a competitor (comp) (in 100× and 200× excess) and in the presence of a mutated WOX4a oligomer (comp^mut) (with 100× and 200× excess)