Fig. 1. Survey of protein kinases for their ability to induce centrosome splitting. U2OS cells were transfected with constructs encoding either wild-type or catalytically inactive protein kinases, as indicated. After 24 hours, centrosomes were analyzed by immunofluorescent staining with antibodies against -tubulin; transfected cells were identified using antibodies against the myc- or HA-tag, as appropriate. (A,B) Cells were classified as harboring either paired (‘un-split’) (A) or split (B) centrosomes, using the criteria described in Materials and Methods. Bar, 10 µm. As illustrated in schematic form, centrosome splitting occurs between the two parental centrioles, each with its associated PCM. Thus, splitting of an S or G₂ phase centrosome produces two products that each comprises one parental centriole and one tightly associated procentriole. The histogram (C) indicates the percentages of cells showing split centrosomes in response to expression of each kinase construct.

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