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. 2019 May 10;10:503. doi: 10.3389/fpls.2019.00503

FIGURE 5.

FIGURE 5

SOG1 directly interacts with KIN10/11 in Y2H and in planta. (A) Yeast two-hybrid (Y2H) assays show the interaction between SOG1 and KIN10, and SOG1 and KIN11. The vectors containing only AD (the activate domain) or BD (the LexA DNA-binding domain) act as negative controls. SOG1, KIN10, and KIN11 bait constructs were fused to the LexA DNA-binding domain and SOG1, KIN10, and KIN11 prey constructs were fused to the activation domain. (B) Schematic diagrams of SOG1 and KIN10/11 domains and constructs used in bi-molecular fluorescence complementation (BiFC). (C–F) BiFC interaction assays in Nicotiana benthamiana pavement cells transiently co-expressing N-terminal yellow fluorescent protein (nYFP)-SOG1 and C-terminal YFP (cYFP)-IN10/11. Panels (C,D) are in normal conditions (treatment with DEX and 2 μg/ml DMSO), and (E,F) are treated with DEX and 20 μg/ml antimycin A. Scale bars: 50 μm. (C–F) Labels above each panel indicate the filter channel imaged. DIC, differential interference contrast images (left side of each panel); YFP, fluorescence images (middle of each panel); and Merge, the DIC and YFP images combined (right side of each panel). SOG1-nYFP/KIN10-cYFP, co-expression of 35S::SOG1-nYFP and 35S:: KIN10-cYFP; SOG1-nYFP/KIN11-cYFP, co-expression of 35S::SOG1-nYFP and 35S::KIN11-cYFP. (E,F) Red arrows indicate nucleus. (G) In vivo interaction of SOG1 with KIN10/11. DMSO acts as a control. Black arrow, SOG1-Myc. IP, immunoprecipitation; WB, Western blotting.