Figure 5.

AtZAR1^CC oligomerizes in yeast and in planta. A, AtZAR1^CC or AtZAR1^CC2 was constructed as a fusion with the DNA-binding domain and the activation domain, and tested for interaction against themselves or empty vector (EV) in the LexA yeast two-hybrid assay. HopF2_PtoDC3000 and its chaperone ShcF2_PtoDC3000 were used as positive controls, because they are known to strongly interact (Shan et al., 2004). The experiment was performed twice with similar results. B, Bimolecular fluorescence complementation assay of AtZAR1^CC interactions. A. tumefaciens carrying AtZAR1^CC-YFP, AtZAR1^CC, and EV as a fusion to the cYFP or nYFP was mixed in equivalent optical densities and pressure-infiltrated into the leaves of N. benthamiana. Left, the YFP channel alone. Right, a merge of the YFP channel and bright field (BF). Leaf sections were imaged using a Zeiss LSM710 confocal scanning microscope 24 to 48 h after induction. The red arrowheads show the nuclei. Bar = 50 μm. The experiment was performed twice with similar results.