FIG. 8.
p55HEF1 associates with the mitotic spindle. Cells in prophase (A to C), metaphase (D to F), late anaphase (G to I), and cytokinesis (J to L) were stained with α-HEF1-R2 antiserum to HEF1 and visualized with rhodamine (A, D, G, and J) or stained with α-tubulin and visualized with fluorescein isothiocyanate (B, E, H, and K); a merged image is shown in panels C, F, I, and L, with HEF1-tubulin colocalization shown in yellow. Note punctate staining of α-HEF1-R2, which does not colocalize with microtubules in nonmitotic cells. Focal adhesion staining is not visible in this optical section, although it is clearly present in nonmitotic cells. Identical results were obtained with α-HEF1-R2 in a single stain, excluding bleedover from tubulin as a source of HEF1 staining (data not shown).