Figure 5.

Interactions between JAZ Repressors and ICE Transcription Factors.

(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.

(B) BiFC analysis. Fluorescence was observed in nuclear compartments of N. benthamiana leaf epidermal cells; the fluorescence resulted from complementation of the N-terminal portion of YFP fused to ICE factors (ICE-nYFP) with the C-terminal portion of YFP fused to JAZ proteins (JAZ-cYFP). No signal was observed from negative controls. DAPI, 4′,6-diamidino-2-phenylindole.

(C) CoIP analysis. MYC-fused JAZ proteins were immunoprecipitated using an anti-MYC antibody, and coimmunoprecipitated ICE factors were then detected using an anti-FLAG rabbit antibody. Protein input for MYC-JAZ proteins in immunoprecipitated complexes was also detected and shown.