Xenbase Image ID: 118868

Figure 2. TACC3 increases the antagonizing activity of XMAP215 against MCAK. (A) Sedimentation analysis to monitor MCAK-dependent microtubule destabilization activity. 2.5 μM GMPCPP microtubules were mixed with 125 nM MCAK (lanes 3–14) or control buffer (lanes 1 and 2) in presence of ATP. 500 nM XMAP215 (lanes 5–14) and increasing concentrations of TACC3 from 250 (lanes 7 and 8), 500 (9 and 10), and 1,000 (11 and 12) to 1,500 nM (13 and 14) or control buffer (5 and 6) were added in reactions. Reaction mixtures were sedimented after a 38-min incubation at 30°C, supernatants (S) and pellets (P) were subjected to SDS-PAGE, and the gel was stained with Coomassie brilliant blue. (B) Sedimentation assay to monitor XMAP215 affinity to microtubules. 2.5 μM GMPCPP microtubules were mixed with 1,250 nM TACC3 (lanes 1 and 2), 500 nM XMAP215 (3 and 4), or 500 nM XMAP215 in addition to increasing concentrations of TACC3 from 250 (5 and 6), 500 (7 and 8), 750 (9 and 10), and 1,000 (11 and 12) to 1,250 nM (13 and 14). Mixtures were incubated for 38 min at 30°C and sedimented, and supernatants (S) and pellets (P) were analyzed by SDS-PAGE. Image published in: Kinoshita K et al. (2005) Copyright © 2005, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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