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. 2018 Jul 4;8:10114. doi: 10.1038/s41598-018-28413-x

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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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Structure of the ABI4 promoter and chromatin immunoprecipitation (ChIP) using a SND1-GFP fusion protein. (A) cis-elements in the ABI4 promoter region. ABRE is an abscisic acid (ABA)-response element; GT1-motif and GAG-motif are light-response elements; CAAT-Box is a common cis-acting element. The PlantCARE program was used for ABI4 promoter analysis (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/)¹⁹. ABI4pro#1, #1–1, and #1–2 show the size and location of the expected PCR products from the DNA template of ChIP. ABI4pro q#1, q#2, q#3, q#4, and q#5 present the size and location of the expected products of qRT-PCR products from the DNA template of ChIP. (B) The fold-enrichment values were calculated by dividing the ChIP signals obtained via the qRT-PCR by the signals obtained for the mock. The error bars indicate the standard error (SE). The asterisks represent significant differences among values (P < 0.05). (C) The yeast one-hybrid assay between SND1 and ABI4 promoter. Full-length SND1 was fused with GAL4 AD, and ABI4pro#1 was used as a bait. The ABI4pro-AbAi/pGADT7 yeast served as the negative control. Yeasts were cultured on SD/-Leu or SD/-Leu with 50 ng/mL aureobasidin A (AbA). (D) The electrophoretic mobility shift assay (EMSA) showed that the SND1 NAC domain (SND1 BD-His MBP) can bind to the biotin-labelled ABI4 promoter q#1.