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. 2022 May 4;13:2438. doi: 10.1038/s41467-022-30254-2

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© The Author(s) 2022

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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Fig. 5 — a Results of yeast two-hybrid assays show BSL2/BSL3/BSU1 proteins directly interact with MPK3/6. Bait, “BD-” indicates Gal4 DNA-binding domain; Prey, “AD-” indicates Gal4 activation domain. “Growth controls”, assays performed using rich media (-Leu-Trp); “Interaction tests”, assays performed using synthetic dropout medium (-Leu-Trp-His; 1 mM 3-AT added to suppress bait auto-activation). b Co-IP assays using purified fusion proteins produced by N. benthamiana leaves show physical association of BSL2-FLAG (left) or BSU1-FLAG (right) with MPK6-YFP. YFP (left lane, - control) or MPK6-YFP (right lane) was used as bait to bind to the GFP-Trap agarose. Immunoprecipitated proteins were detected by anti-FLAG. Data represent results of three biological repeats. c Results of bimolecular fluorescence complementation (BiFC) assays in N. benthamiana leaf epidermal cells show BSL2 and BSU1 interact with MPK6 in plant cells. YFP signals (green) indicate protein-protein interactions. No interactions were detected between BSL2/BSU1 and the transcription factors SPCH/SCRM. nYFP, N-terminal YFP; cYFP, C-terminal YFP. Data represent results of three independent experiments. Scale, 50 μm. (z), z-staked confocal images.