(
A) Background-subtracted GFP intensity plotted against the distance between separating chromosome masses at 7 min to assess dependence of recongression on expression level (Spearman correlation coefficient –0.6246, one-tailed p-value<0.0001). Gray dashed line indicates mean background subtracted GFP intensity of 100. Forty-three KIF22-GFP cells from 10 experiments, 21 KIF22-GFP P148L cells from 6 experiments, 28 KIF22-GFP P148S cells from 7 experiments, 16 KIF22-GFP R149L cells from 6 experiments, 17 KIF22-GFP R149Q cells from 4 experiments, and 21 KIF22-GFP V475G cells from 21 experiments. (
B) Distance between separating chromosome masses of cells expressing lower levels of KIF22-GFP (mean background subtracted GFP intensity less than 100). Lines represent the mean and the shaded area denotes SEM. Twenty-seven KIF22-GFP cells from nine experiments, 16 KIF22-GFP P148L cells from six experiments, 18 KIF22-GFP P148S cells from six experiments, 16 KIF22-GFP R149L cells from six experiments, 6 KIF22-GFP R149Q cells from three experiments, and 7 KIF22-GFP V475G cells from three experiments. (
C) Distance between separating chromosome masses 7 min after anaphase onset of cells expressing lower levels of KIF22-GFP (mean background subtracted GFP intensity less than 100). Bars indicate medians. P values from Kruskal-Wallis test. P values are greater than 0.05 for comparisons without a marked p value. Data represent the same cell populations presented in (
B). (
D) Time-lapse images of dividing RPE-1 cells expressing KIF22-GFP R149Q or KIF22-GFP V475G. Imaged approximately 12–18 hr after treatment with doxycycline to induce expression. Times indicate minutes after anaphase onset. Images are maximum intensity projections in z through the entirety of the spindle. Scale bar 5 μm. Images are representative of three or more experiments. (
E) Distance between separating chromosome masses throughout anaphase in RPE-1 cells. Lines represent the mean and the shaded area denotes SEM. Twenty-five KIF22-GFP cells from seven experiments, 16 KIF22-GFP R149Q cells from six experiments, and 8 KIF22-GFP V475G cells from six experiments. (
F) Distance between separating chromosome masses 7 min after anaphase onset in RPE-1 cells. Bars indicate medians. P value from Kruskal-Wallis test. P values are greater than 0.05 for comparisons without a marked p value. Data represent the same cell populations presented in (
E). See
Figure 4—figure supplement 1—source data 1.