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Fig. 5. Cdc2 and Aurora-A kinase activation is independent of PP1 inhibition. Prophase extracts were depleted (right panels) or not (left panels) in PP2A by incubation at 4°C with microcystin-agarose beads. The extracts were then incubated in the presence or absence of 500 nM P-DARPP-32 at 30°C with an ATP-regenerating system (ATP) plus or minus okadaic acid (OA). Samples were collected at indicated times for histone H1 (A) and histone H3 kinase assays (B). (A) Samples were pulled down on p13 beads and assayed for Cdc2 kinase activity using histone H1 as substrate. (B) Samples were immunoprecipitated with anti-Aurora-A antibody and assayed for Aurora-A kinase activity using histone H3-peptide as substrate. Cdc2 and Aurora-A kinase activities are expressed in cpm incorporated in the substrate.





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