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. 2016 Apr 20;7:530. doi: 10.3389/fpls.2016.00530

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Copyright © 2016 Kwak, Son, Kim, Song and Seo.

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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Interaction of HPY2 with FLC. (A) His₆-FLC and GST-HPY2 were overexpressed in Escherichia coli and purified with Ni²⁺-NTA or glutathione affinity columns. (B) The His₆-FLC protein was pulled down with the GST-HPY2 protein, separated on 11% SDS-polyacrylamide gels, and analyzed by Western blotting with anti-His antibody. (C) In vivo interaction of HPY2 and FLC. Wild-type (WT) plants were infiltrated with different combinations of 35S-Myc₆-FLC or 35S-HPY2-FLAG₃ constructs. Total protein was extracted from each sample and immunoprecipitated with anti-FLAG antibody. After immunoprecipitation, Myc₆-FLC was detected by Western blotting with anti-Myc antibody. Myc₆-FLC and HPY2-FLAG₃ expression was also examined by Western blotting with anti-Myc and anti-FLAG antibodies, respectively.