Figure 2. The C-terminal domain of Ska3 is required for timely mitotic progression.

(a) Quantification of MT-cosedimentation assays comparing the wt Ska complex and Ska1-Ska2-Ska3ΔC with Ska1-Ska2-Ska3_1–151 and Ska1-Ska2-Ska3_1–195. K_d values were calculated using 1 μM Ska and 0–12 μM MTs (mean ± s.d., n = 4). (b) Box-and-whisker plot showing the elapsed time (min) between nuclear envelope breakdown (NEBD) and anaphase onset/death for individual cells. The number of cells (n) from three independent time-lapse experiments is given above each box. Lower and upper whiskers represent 10^th and 90^th percentiles, respectively. Below, table summarizing information from the live cell experiments regarding the average time between NEBD to anaphase onset/death and the percentage of cells dying in mitosis. Myc-V and mCherry-V were used as transfection controls. (c) Representative stills from time-lapse video-microscopy experiments illustrating mitotic progression of HeLa S3 cells stably expressing histone H2B-GFP treated as in (b). Time in hr:min is indicated. T = 0 was defined as the time point where NEBD became evident. Scale bar, 10 μm.