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Figure S2. A time-lapse sequence of an^{early mitotic LLC-PK cell expressing full-length Cdc20-GFP showing both the fluorescence andphase contrastimages.The numbersindicate time inminutesbefore andafter NEB.Initial accumulationof Cdc20-GFP atkinetochoreswasdetected3 minbefore NEB (-3 min, white arrow).All the kinetochoresshowedbrightCdc20-GFP fluorescence <1 minbefore NEB (white arrows).The spindle polesare markedwith arrowheadswheninthe image focal plane.Notable differenceswere observedinthe intensity of the kinetochore-boundCdc20-GFP signal betweenchromosomesnear the poles(white arrowheads) andchromosomesalignedatthe metaphase plate (black arrowheads).However, no differenceswere seenbetweenthe leading andtrailing kinetochoresof individual unalignedor alignedchromosomes(white linesin+8 and+11 min).A Quicktime video of the time-lapse phase andGFP image (Video 1) of thiscell isalso available.}