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. 2021 Jan 16;22(2):870. doi: 10.3390/ijms22020870

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Identification of interactions between JAZs and bHLHs. (A) Y2H assay to screen the interaction between all JAZs and bHLH99, bHLH91 and MYC7 proteins. bHLH91.1 represents the N-terminus of bHLH91 while bHLH91.2 represents C-terminus. MYC7.1 represents the N-terminus of MYC7 and MYC7.2 for C-terminus. (B) Split-luciferase assay confirming the interactions between selected JAZs and MYC7 (top panels) and bHLH99 (bottom panels). Panel pictures in a, b and c represent different JAZ members to be tested with either MYC7 or bHLH99, and d is the schematic diagram for different combination of the vectors. (C), Pull-down assay confirming the interaction between JAZ15 and MYC7. The assay was conducted using 2 µg COI1-MBP and JAZ-GST fusion protein each reaction in the presence of 20 µL GST beads. The bands marked in red triangle in top panel shows the signal identified by MBP antibody, and the bands marked in red star in bottom panel shows the input of JAZ-GST fusion protein and GST protein using GST antibody.