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. 2002 Aug;14(8):1847–1857. doi: 10.1105/tpc.002550

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Copyright © 2002, American Society of Plant Biologists

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Figure 3. — Kinase Activity of Mutant Nicta; CycD3;3 in Vitro.

(A) Lysates from insect cells expressing the indicated proteins were assayed for the phosphorylation of histone H1 and NtRb1. Labeled histone H1 and NtRb1 were resolved on denaturing polyacrylamide gels and visualized by autoradiography.

(B) Mutant Nicta; CycD3;3 forms a complex with Nicta; CDKA;3. Lysates from insect cells expressing FLAG-tagged Nicta; CDKA;3 (lanes 3, 4, 7, and 8) and either His-tagged wild-type Nicta; CycD3;3 (lanes 1 to 4) or His-tagged mutant Nicta; CycD3;3 (T191) (lanes 5 to 8) were purified on an anti-FLAG M2 affinity gel. Crude extracts (C) and elute fractions (E) were separated on 10% SDS–polyacrylamide gels. FLAG-tagged Nicta; CDKA;3 was detected by immunoblotting with an anti-FLAG M2 monoclonal antibody (bottom gel); His-tagged wild-type or mutant Nicta; CycD3;3 were detected by immunoblotting with a His tag–specific antibody (top gel).