Figure 6.

Protein binding profile of the Znf-BED domain of Tam3 TPase. A, Mass protoplasts prepared from young Antirrhinum leaves transformed with Tam3 TPase constructs wild-type (wt) and m10 (see Fig. 5). The protoplasts with wild type and m10 showed GFP expression in the PM and nucleus, respectively. Transformation efficiencies for wild-type and m10 constructs were 40 to 50%. Protein preparations were made using these Antirrhinum protoplasts. B, CBB staining of the native PAGE gel for nondenatured proteins extracted from the three protoplast samples, nontransformant protoplasts (control), transiently transformed protoplasts with wild-type construct and m10 construct (m10). Active proteins were extracted using native protein extraction buffer containing 0.20% Triton X-100. C, GFP-Tam3 TPase complex detected by western blotting using an anti-GFP antibody. The duplicate of the gel blot in B was made by loading adjacent lanes with the same samples. Gray arrowhead indicates specific band to wild-type samples, and white arrowhead shows band shared by wild-type and m10 samples.