Supplemental Material

This article contains the following supporting material:

• Supplementary Figure 1. - An increased amount of MCAK on chromosome arms in hesperadin-treated extracts is MCAK specific.
  Chromatin assembled in CSF egg extracts in the presence of control DMSO (Control) or 5 μM hesperadin (Hesp), pelleted onto coverslips, and immunostained with α-MCAK, α-Ndc80, α-CENP E or α-P150. Scale bar, 5 μm.
• Supplementary Figure 2. - Cytoplasmic MT assembly is not completely inhibited in the absence of Aurora B activity.
  (A) MT asters were induced in Xenopus egg extracts containing rhodamine-labeled tubulin with 25 μM of His-RanL43E (top) or 5% DMSO (bottom) in the absence (Control) or presence of 5 μM hesperadin (Hesp). Reactions were incubated for 20 minutes (for Ran asters) or 30 minutes (for DMSO asters), pelleted onto coverslips, and mounted in mounting medium. (B) Sperm nuclei were added to extracts containing rhodamine-labeled tubulin to induce spindle formation in the absence (Control) or presence of 5 μM hesperadin (Hesp). Reactions were incubated for 10 minutes (top) or 30 minutes (bottom), pelleted onto coverslips, and mounted with mounting medium. MTs are in red and DNA is in blue. Scale bar, 10 μm.
• Supplementary Figure 3. - MCAK-2-149(T95A) does not target to centromeres in the absence of Aurora B activity.
  Chromatin was assembled in CSF egg extracts treated with control DMSO (Control) or 5 μM hesperadin (Hesp) in the presence of MCAK-2-149 or MCAK-2-149(T95A). Chromatin was pelleted onto coverslips and immnostained α-GST antibodies to show the localization of MCAK-2-149 and MCAK-2-149(T95A).