Figure 3.

Time-lapse imaging of Klp67A-GFP and Klp10A-GFP during mitosis. (A) Dynamics of Klp67A-GFP from metaphase to late anaphase. Clear punctate signals are visible at kinetochores and fainter signals are along K-fibers. Kinetochore signals (yellow arrowheads) are clearly seen during metaphase and diminish during anaphase A (90–150 s). Central spindle accumulation (blue arrowhead) begins at the onset of anaphase spindle elongation. Bar, 5 μm. See also Movie 1. (B) Dynamics of Klp10A-GFP from prophase to prometaphase. Clear punctate signals are visible on the astral microtubules emanating from centrosomes (red) and at centromeres (yellow) during prophase, whereas astral microtubule staining was dramatically reduced after NEB (340 s). Aggregated GFP signals are temporarily visible in cytoplasm, the function of which is unclear. Bar, 10 μm. See also Movie 2. (C) Dynamics of Klp10A-GFP from metaphase to late anaphase. Clear signals are visible at centrosomes (red), pole regions where minus ends of K-fibers are focused (blue) and at centromeres. Centromere signals are seen during metaphase and persist until the end of anaphase A (yellow). Nearly uniform spindle localization is detected during late anaphase spindle elongation. Bar, 10 μm. See also Movie 3.