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. 2018 Oct 18;175(3):796–808.e14. doi: 10.1016/j.cell.2018.09.029

Figure S2.

Figure S2

KIF11-Dependent Organization of Nematic Networks of Extensile Bundles, Related to Figure 1

(A) Binary confocal fluorescence microscopy images showing at high time-resolution the time course of dynamic fluorescent microtubule bundle extension within a nematic network organized by KIF11. (B) Binary confocal fluorescence microscopy images showing the separation of two photo-bleached marks (yellow asterisks) in an extensile bundle within a nematic network organized by KIF11. Binary images (after background subtraction and thresholding) of the microtubule channel are presented to enhance the visibility of distinct network parts. Time is in min:s. (C) Nematic networks organized by KIF11 are three-dimensional as revealed by confocal imaging of different focal planes in the flow chamber. Experiments presented in A - C were performed in the presence of 27 nM KIF11-mGFP, 30 μM tubulin, and 1000 nM mCherry-CAMSAP3-C. (D) Scatterplot depicting microtubule growth episode lengths at different CAMSAP3-C concentrations. Number of growth episodes measured at different mGFP-CAMSAP3-C concentrations: 250 nM – 80, 500 nM – 54, 1000 nM – 107. Horizontal lines indicate the mean and the standard deviation. (E) Box-and-whiskers plot depicting the dependence of KIF11-driven microtubule gliding speeds on the KIF11-mGFP concentration used to immobilise the motor on the glass surface for gliding assays with GMPCPP-stabilized microtubules. The measured speeds agree with previously reported speeds of metazoan kinesin-5 motors (Cole et al., 1994, Hentrich and Surrey, 2010, Kapitein et al., 2005, Krzysiak et al., 2006, Ma et al., 2011, Sawin et al., 1992, van den Wildenberg et al., 2008). The boxes extend from 25th to 75th percentiles, the whiskers extend from 5th to 95th percentiles, and the mean value is plotted as a line in the middle of the box. Number of gliding episodes measured at different KIF11-mGFP concentrations: 20.5 nM – 77, 41 nM – 87, 82 nM – 150, 164 nM – 128, 328 nM – 105. All experiments were carried out in self-organization buffer at 33°C.