Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Mar 15;15(3):e1007993. doi: 10.1371/journal.pgen.1007993

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2019 Zheng et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PMC Copyright notice

Fig 3 — (A) BiFC assays showing that HAT1 interferes the interaction of TT8 or EGL3 with MYB75. YFP fluorescence was observed 48 h after coexpression of TT8-nYFP or EGL3-nYFP/MYB75-cYFP (Control), HAT1-Flag/TT8-nYFP or EGL3-nYFP/MYB75-cYFP (+HAT1-Flag), or Flag/TT8-nYFP or EGL3-nYFP/MYB75-cYFP (+Vector). Bar = 100 μm. (B-C) Quantification of YFP fluorescence intensity in (A). Fifty separate fluorescent flecks were evaluated for fluorescence intensity. Error bars denote ± SD (n = 3). (D-E) qPCR analysis of HAT1, TT8-nYFP (TT8), EGL3-nYFP (EGL3) and MYB75-cYFP (MYB75) expression in (A). The primers used for amplificating Arabidopsis TT8-nYFP (TT8), EGL3-nYFP (EGL3) and MYB75-cYFP (MYB75) are shown in S1 Table online. Expression levels were standardized to Nicotiana Actin, and results of control treatment were set at 1. Error bars denote ± SD (n = 3). (F) Competitive binding assays of TT8 or EGL3 and HAT1 to MYB75. The gradient indicates the increasing amount of HIS-HAT1. The immunoprecipitated fractions were detected by anti-MBP antibody. HIS-HAT1 input is shown in the lower panel. (G) The effects of HAT1 on the MYB75-TT8 and MYB75-EGL3 interaction in plant cells. HA-MYB75 was expressed in protoplasts from wild-type or transgenic plants overexpressing HAT1-GFP with MBP-TT8 or MBP-EGL3. The interaction was detected by immunoprecipitation on anti-MBP resin. The immunoprecipitated proteins (IP) were detected with an antibody recognizing HA. The expression levels of HA-MYB75 and HAT1-GFP were also detected using the indicated antibodies. (H) HAT1 competes with TT8 or EGL3 for binding to MYB75 in yeast. Modulated by the Methionine (Met)-suppressive MET25 promoter, HAT1 was co-expressed with indicated AD and pBridge fusion proteins. Yeast cells grown on the media suppied with the indicated Met levels.