Figure 5.

Interaction between WRKY75 and DELLAs. A, Yeast two-hybrid assay analysis. Interaction was indicated by the ability of cells to grow on synthetic dropout medium lacking Leu, Trp, His, and Ade. The GAL4 activation domain expressed by pGADT7 (shown as “AD”) was used as negative controls. The experiment was repeated three times with similar results and representative photos were displayed. B, CoIP analysis. GFP-fused RGL1 and GAI were immunoprecipitated using anti-GFP antibody, and coimmunoprecipitated Myc-WRKY75 was then detected using anti-Myc antibody. Protein input for GFP-RGL1 and GFP-GAI in immunoprecipitated complexes were also detected and are shown. The experiment was repeated three times with similar results and representative photos were displayed. C, BiFC analysis. Fluorescence was observed in nuclear compartments of N. benthamiana leaf epidermal cells; the fluorescence resulted from complementation of the C-terminal portion of YFP fused to RGL1 or GAI (RGL1-cYFP and GAI-cYFP) with the N-terminal portion of YFP fused to WRKY75 (WRKY75-nYFP). No signal was observed from negative controls. The experiment was repeated three times with similar results and representative photos were displayed. Scale bar: 25 μm. DAPI, 49,6-diamidino-2-phenylindole.