Depletion of Cdt1 from mitotic cells does not interfere with the normal targeting of the Ska1 complex. (A) Schematic representation of the knockdown protocol in double thymidine synchronized HeLa cells that are used to deplete Cdt1 specifically during mitosis. (B) Representative Western blots shown with indicated antibodies in each case to assess the level of Ska3 knockdown upon siRNA addition. (C) Same as in B but to assess the level of Cdt1 knockdown upon siRNA addition. (D) HeLa cells treated with either scramble control or siRNA against endogenous Cdt1 were fixed using paraformaldehyde. Representative images of cells immunostained with antibodies against Hec1 (kinetochore marker) in red and SKA3 (representing the Ska1 complex) in green are shown. Scale bar, 5 μm. (E) HeLa cells treated with either scramble control or siRNA against endogenous Cdt1 were fixed using paraformaldehyde. Representative images of cells immunostained with antibodies against SKA3 (in green) and tubulin (in red) are shown; DAPI stained the chromosomes and shown in black and white. Scale bar, 5 μm. Source data are available for this figure: SourceData FS2.