Fig. 7. Eg5 and AtKRP125c in fixed animal epithelial cells. (A,B) Interphase. AtKRP125c-myc localized to microtubules in LLC-Pk1 cells at interphase, whereas Eg5 did not. (C,D) Metaphase. Both AtKRP125c-myc and Eg5 were strongly localized to the spindle in mitotic cells. (E) Treatment of LLC-Pk1 cells with monastrol caused the formation of monopolar spindles and cell cycle arrest, which was not affected by the presence of AtKRP125c-myc. (F,G) Mitotic LLC-Pk1 cells transfected with a hairpin construct against Eg5 were clearly visible by the presence of monopolar spindles and diminished Eg5 labeling (F, arrowheads), whereas cells that were not knocked down had bipolar spindles that labeled strongly for Eg5 (F, arrows). In cells co-transfected with the Eg5 hairpin construct and the AtKRP125c-Myc construct (G), the great majority of spindles were monopolar, despite AtKRP125c binding to spindle microtubules. Bar, 10 µm.