SKAP ΔEB mutant cells display a spindle mispositioning defect. (A, left) IF images showing microtubules in the SKAP ΔEB mutant spindle. Images represent maximum-intensity projections. The dashed line indicates the cell boundary. (right) Graph showing difference in the distances between each spindle pole and the closest position on the cell cortex (see diagram at bottom right of the figure). A value of 0 represents spindle positioning in the cell center, with equivalent distances to each cell cortex. n = 100 total cells per condition collected from two (ΔEB without depletion) or three independent experiments. Mean and SD are plotted. ****, P < 0.0001, significant difference assessed by an unpaired two-tailed t test. *, difference with P = 0.0153. (B, left) Images showing maximum-intensity projections for microtubule staining to show for spindle mispositioning phenotype and its suppression by LGN depletion. (right) Graph showing the pole–cortex difference for LGN experiment (plotted as in A). n = 100 cells per condition collected from three independent experiments. ****, P < 0.0001, significant difference assessed by an unpaired two-tailed t test. (C, left) Maximum-intensity projections for microtubule staining show the spindle mispositioning phenotype and its suppression by 20 nM nocodazole. (right) Graph showing the pole–cortex difference (as in A and B) for the low-dose nocodazole experiment. n = 100 cells per condition collected from two independent experiments. Mean and SD are plotted. ****, P < 0.0001, significant difference assessed by an unpaired two-tailed t test. (D) Kymographs of videos from cells in which wild-type (WT) SKAP or the SKAP ΔEB mutant replaces endogenous SKAP. Arrows mark the start of the spindle shift in ΔEB cells. Also see Video 4. DIC, differential interference contrast.