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. 2018 Aug 9;8:11929. doi: 10.1038/s41598-018-30166-6

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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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Chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) analysis of the DNA binding activity of mTERF6 in vivo. (A) Immunoprecipitation and immunoblot analysis of mTERF6. The samples consisting of the total leaf protein extract from pMTERF6::MTERF6.1-FLAG transgenic plants (mterf6-5 background) were immunoprecipitated with the FLAG antibody-conjugated protein G beads (IP+) or naked protein G beads (IP−) as a control. The immunoblot analysis was performed with the FLAG antibody. The lane “mTERF6.1-FLAG” is the input. (B) The position of the designed primer pairs for ChIP-qPCR analysis. (C) DNA immunoprecipitation analysis. The chloroplasts fixed with formaldehyde (FA) were extracted from the pMTERF6::MTERF6.1-FLAG transgenic plants, then all of the DNA was sonicated into short fragments under specific conditions and incubated with the FLAG antibody-conjugated protein G beads. With the interaction between the FLAG antibody and the mTERF6.1-FLAG protein, the associated DNA was immunoprecipitated and analyzed by quantitative real-time RT-PCR. Error bars indicate standard deviations for triplicates.