flp1p is phosphorylated during mitosis.

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flp1p is phosphorylated during mitosis. flp1p is phosphorylated during mitosis. The strain cdc25-22 flp1-HAc was synchronised by arrest-release and protein samples were prepared from cells at the indicated times. Note that a slower-migrating form of flp1p-HA appears during mitosis. The black arrowhead (B) indicates the rapidly migrating, interphase form of the protein, the grey arrowhead indicates the form present in G2 cells. (A) The progression through mitosis after release from the cdc25-22 block, as determined by fixing cells at each time point, and staining with DAPI and Calcofluor. (B) Western blot of protein samples prepared at the indicated times, probed with 12CA5 and TAT-1, to reveal flp1p-HA and α-tubulin, respectively. (C) flp1p-HA was immunoprecipitated from protein samples prepared at the indicated times. After washing, the 45 minute immunoprecipitate was divided in half and treated with alkaline phosphatase (+), or not (-). A western blot of these protein samples was probed with 12CA5 to reveal the position of flp1p-HA. Nathalie Cueille et al. J Cell Sci 2001;114:2649-2664 © The Company of Biologists Limited 2001