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Fig. 2. The BRCT domain targets Ect2 to the central spindle, whereas the PH domain is required for cell cortex localization. (A) Schematic representation of different Ect2 domains and the ability of various N- and C-terminal fragments to localize to the central spindle and the cell cortex. + indicates presence and – absence of localization to the central spindle and cortex. (B) HeLa S3 cells were grown on coverslips and transiently transfected for 36 hours with the indicated myc-Ect2 fragments before fixation with paraformaldehyde and permeabilization with Triton X-100. Cells were stained with anti-myc 9E10 antibodies (left) and FITC-phalloidin (middle). Merged images are shown on the right with 9E10 in red, actin in green and DNA stained with DAPI in blue. (C) HeLa S3 cells were transfected for 36 hours with the indicated myc-Ect2 constructs, fixed and permeabilized as described in B and stained with anti-myc 9E10 antibodies (left) and anti -tubulin antibodies (middle). Merged images are shown on the right, with 9E10 in red, -tubulin in green and DNA stained with DAPI in blue. Bars, 10 µm.

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