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. 2016 Nov 22;7:1728. doi: 10.3389/fpls.2016.01728

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Copyright © 2016 Meng, Li, Liu and Jiang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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AN3 negatively regulates EMB71/YDA expression. (A) Bar graph exhibiting the difference of expression of EMB71/YDA between at least 10 Wild-type (Col-0), an3-1, an3-4, and 35S:AN3/an3-4 seedlings grown under white light conditions. (B) Bar graph exhibiting the difference of expression of AN3 between Wild-type (Col-0), emb71, Ler, yda-1, and yda-2 seedlings grown under white light conditions; Data were from quantitative RT-PCR. Materials were from seedlings of at least five independently propagated lines. And wild-type is set as 1.0. Quantifications were normalized to the expression of UBQ5. Error bars represent SD (n = 3). Heteroscedastic t-test analysis showed significant differences (^∗∗∗P < 0.001). In (A,B) these experiments were repeated at least two times with similar results (technical replicates). (C) A chromatin immunoprecipitation (ChIP) analysis. Enrichment of particular YDA chromatin regions with anti-HA antibody (as a control) or anti-GFP antibody in 35S:AN3-3XGFP transgenic plants as detected by real-time PCR analysis. (D) A chromatin immunoprecipitation (ChIP) analysis. Enrichment of particular YDA chromatin regions with anti-GFP antibody (as a control) or anti-HA antibody in 35S:AN3-HA transgenic plants as detected by real-time PCR analysis. Quantifications were normalized to the expression of UBQ5. Error bars represent SD (n = 3). Heteroscedastic t-test analysis showed significant differences (^∗∗P < 0.01). Input is set as 100% [supernatant including chromatin (input material) is considered as 100%, immunoprecipitated chromatin/input material X 100% for enrichment product of particular YDA chromatin regions]. In (C,D) these experiments were repeated at least two times with similar results (biological replicates). (E) Schematic of the YDA promoter loci and their amplicons for ChIP analysis. (F) ProYDA:GFP in WT (Col-0) (a) and 35S:AN3 (b) cotyledons. they are at same magnification. White arrows point to GFP-positive nuclei.