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Cdc28 and Kin28 are phosphorylated in vivo. HA epitope-tagged Cdc28, Kin28, and Pho85, under the control of their own promoters, were immunoprecipitated from cell lysates with the 12CA5 antibody and treated with phosphatase buffer alone (lanes 1, 4, and 7), λ-phosphatase (λ PP’ase) (lanes 2, 5, and 8), or λ-phosphatase in the presence of phosphatase inhibitors (lanes 3, 6, and 9). The immunoprecipitates were then subjected to immunoblotting with 12CA5.
