Spindle misorientation induced by hSpindly depletion depends on dynein. (A) Representative stills from videos of GFP-α-tubulin/cherry-H2B expressing HeLa Kyoto cells treated with the indicated combinations of siRNAs for 54 h and synchronized by double thymidine block and release. 9 h after the second release, MG132 was added for 1 h before filming. Time is shown in h:min. t = 0 was defined as the time point where chromosomes aligned in the metaphase plate. (B) Bar graphs showing the percentages of cells with spindle rotation defects. Error bars show the SD from three experiments (>15 cells per experiment). (C) Cells stained with anti-p150^Glued (green) and anti-hSpindy antibodies (red), and DAPI (blue). Arrows indicate p150^Glued cortical staining. (D) Cells were treated with GL2 or hSpindly siRNAs for 48 h and stained with anti-p150^Glued antibody and DAPI. Arrows indicate p150^Glued cortical staining. Bars, 10 µm.