(A) Kymograph of the displacement of 230 pM Eg5-513-GFP dimers versus time in 80 mM Pipes buffer with 0

2 mM ATP. A large majority of binding events last two frames or less (<p><b>Copyright information:</b></p><p>Taken from "Microtubule cross-linking triggers the directional motility of kinesin-5"</p><p></p><p>The Journal of Cell Biology 2008;182(3):421-428.</p><p>Published online 11 Aug 2008</p><p>PMCID:PMC2500128.</p><p></p

(A and B) Frames (A) and kymograph (B) from a time-lapse recording showing single molecules of Eg5-GFP (green) moving directionally along a microtubule (red) in the presence of 70 mM Pipes

(C and D) Frames (C) and kymograph (D) from a time-lapse recording showing single molecules of Eg5-GFP (green) diffusing along a microtubule (red) in the presence of 70 mM Pipes plus 40 mM KCl. (E–H) MD calculated from Eg5-GFP motility in the presence of ATP (black) and ADP (red) in 70 mM Pipes plus 0, 20, 40, or 60 mM KCl. Fits represent MD = . (I–M) MSD calculated from Eg5-GFP motility in the presence of ATP (black) and ADP (red) and at the indicated ionic strengths. Fits represent MD = and MD 2 + offset for the ATP data and MD 2 + offset for the ADP data. All numerical results are listed in . (N) Histogram of the duration of binding events for 0 mM KCl and for 60 mM KCl added. Lines are single exponential fits (exp[−t/t]) to the data (0 mM: t = 34 ± 3, = 212; 60 mM: t = 16 ± 2, = 119). Error bars represent SD. Bars, 1 μm.<p><b>Copyright information:</b></p><p>Taken from "Microtubule cross-linking triggers the directional motility of kinesin-5"</p><p></p><p>The Journal of Cell Biology 2008;182(3):421-428.</p><p>Published online 11 Aug 2008</p><p>PMCID:PMC2500128.</p><p></p

Data collected as described in , but in the presence of a mixture of 1 nM Eg5-GFP and 2 nM of unlabeled tetrameric Eg5

(A) Top kymograph shows sliding of a microtubule (MT) relative to a surface-attached microtubule. Below, the corresponding kymograph of Eg5-GFP shows directional runs between the overlapping microtubules (region marked with two red dotted lines). (B–F) Analysis of Eg5 motility during relative sliding. (B) Scatter plot of all pairs of short-term velocity and diffusion constant determined for a window of 15 s moving over the composite position-time trace of 94 Eg5 motors traced in the overlap zone of 11 microtubule pairs (2,335 points obtained from 2,349 s of total time). The horizontal dotted line indicates the average velocity of sliding microtubules (33 nm/s), and the vertical dotted line indicates the threshold used to discriminate slow and fast diffusion. (C and D) Similar analyses for Eg5 moving on individual microtubules at low ionic strength (C, 70 mM Pipes; ; 4,266 points) and high ionic strength (D, data pooled from 70 mM Pipes + 60 mM KCl and 70 mM Pipes + 80 mM KCl; 2,478 points). (E) Position-time traces. Black, fraction of the composite trace used for B. Green and red, sorted time points with a short-term diffusion constant; < 1,500 nm/s (green) and >1,500 nm/s (red). (F) Histograms of the short-term velocities as obtained from the time points in the green and red trace in E. The arrow indicates the average microtubule sliding velocity. (G) Graph summarizing Eg5 behavior under various conditions. Bar, 2 μm.<p><b>Copyright information:</b></p><p>Taken from "Microtubule cross-linking triggers the directional motility of kinesin-5"</p><p></p><p>The Journal of Cell Biology 2008;182(3):421-428.</p><p>Published online 11 Aug 2008</p><p>PMCID:PMC2500128.</p><p></p