Phosphoregulation of metaphase chromosome oscillations. (A) Kymographs (time and distance scale bars are 24 s and 2 µm, respectively) of representative sister kinetochore pairs from bioriented chromosomes in cells rescued with WT Hec1-GFP or the indicated phosphomimetic mutants generated in an A background. (B) Representative tracings for two sister kinetochore pairs from the indicated backgrounds. The y axis shows the relative position along the spindle axis; time and distance scale bars are the same for all graphs. The vertical offset between each set of pairs is for easier visualization. (C) Mean velocity of kinetochore movement along the spindle axis. See legend on the right for color coding. Data for each bar in C–E are based on 22 kinetochore tracks from at least seven cells. Here and in D and E, the results for 1D Hec1 are the means for three different mutants (Fig. S2, A–C). (D) Time spent with no motion for two sequential frames, or 6 s, normalized to the total time of the time lapse. (E) Deviation from average position, a measure of oscillation amplitude (Stumpff et al., 2008). Error bars are SEMs.