SKAP localizes to the outer kinetochore during metaphase and anaphase. (A) Images of different mitotic stages from a clonal human cell line stably expressing moderate amounts of GFP^LAP-SKAP. GFP-SKAP localizes to spindle microtubules throughout mitosis, and kinetochores during metaphase and anaphase. (B) Immunofluorescence showing the colocalization of SKAP and Hec1/Ndc80 with DNA (blue) and CENP-A (red). (C, left) Immunofluorescence image showing the colocalization of SKAP (green), Hec1 (blue), and CENP-A (red). (C, right) Graph showing a line scan from the boxed kinetochore pair showing the relative spatial distribution of SKAP, Hec1, and CENP-A. SKAP localizes slightly peripherally to Hec1 at the outer kinetochore. (D) Localization of chicken ggSKAP in DT40 cells. (D, top) SKAP alone. (D, bottom) Colocalization of DNA (blue) and CENP-T (red). Insets show enlarged views of the boxed regions. Bars, 5 µm.