Figure 4.

The N-terminal domain regulates physiological MT dynamics and the C-terminal domain regulates spindle bipolarity. Asters and spindles were induced by the addition of 25 μM RanL43E to control or MCAK-depleted extracts to which 200 nM GFP, GMCAK, GM(187-731), GM(2-592), GM(187-582), GM(R522A), GM(E529A), and GM(296-308Δ) were added. Reactions were incubated for 45 min at room temperature, fixed, sedimented onto coverslips, and mounted with anti-fade. (A) Representative images are shown and were scaled identically. Scale bar, 10 μm. (B) Quantification of the structure type is graphed as the mean percentage of the total structures plus the SEM from three independent experiments. (C) Quantification of the aster types is graphed as the mean percentage from the aster and spindle total plus the SEM from three independent experiments. Small asters and spindles, which represent rescued structures, were combined and are compared with large asters. An asterisk indicates significant difference relative to GMCAK add-back.