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Fig. 6. Phosphorylation of cyclin D3 by GSK-3ß. (A) Bacterially produced His-cyclin D3 (wt) (4 µg) was incubated with increasing concentrations of purified GSK-3ß in the presence of [{gamma}-32P]ATP for 30 minutes at 30°C. Following incubation, the samples were subjected to SDS-PAGE and autoradiography. One representative experiment of three is shown. (B) Bacterially produced His-cyclin D3 (wt) (4 µg), His-cyclin D3 (T283) (4 µg), and purified GSK-3ß (300 mU) were incubated in various combinations as indicated with [{gamma}-32P]ATP in the presence or absence of 50 mM LiCl for 30 minutes at 30°C. The reactions were subjected to SDS-PAGE and autoradiography. One representative experiment of three is shown.





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