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Figure 2


Fig. 2. DLG1 is phosphorylated by CDK1 and CDK2 in vitro. (A) Schematic showing the various deletion mutants used to map the regions that are phosphorylated by CDK1 and CDK2. CT, C-terminus; GUK, guanylate kinase domain; NT, N-terminus. (B) GST-DLG1 was incubated with purified CDK1 and CDK2 plus radiolabelled ATP for 20 minutes, and phosphorylation of DLG1 ascertained by SDS-PAGE and autoradiography (upper panel). The lower panel shows the Coomassie-blue-stained gels showing the control GST and the GST-DLG1 fusion protein; the arrow indicates the expected position of the non-phosphorylated GST. (C,D) GST-DLG1 and its various GST-tagged deletion mutants (Fig. 2A) were subjected to in vitro kinase assays with CDK1 and CDK2, respectively. Phosphorylation is observed in the wild type, the N-terminus and N-terminus+3 PDZ regions, but not in the C-terminal region.





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