Figure 3.
Tubulin and KIF11 Concentrations Influence Microtubule Network Organization
(A) Box-and-whiskers plot depicting microtubule plus-end growth speeds at different CAMSAP3-C concentrations. Number of plus-end growth episodes measured at different tubulin concentrations: 7.5 μM, 31; 15 μM, 17; 30 μM, 72. 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. The same data is plotted for the 30 μM condition at 500 nM mGFP-CAMSAP3-C as for Figure 2A.
(B) TIRF microscopy images of CF640R-labeled microtubules showing enhanced microtubule formation at increasing tubulin concentrations in the presence of 500 nM mGFP-CAMSAP3-C (imaged 2 min 20 s after initiating microtubule nucleation).
(C–F) Confocal fluorescence microscopy images showing the time course of KIF11-mediated organization of different types of networks at the following respective concentrations for tubulin and KIF11: 20 μM and 27 nM (C), 10 μM and 27 nM (D), 7.5 μM and 82 nM (E), and 15 μM and 164 nM (F). The mCherry-CAMSAP3-C concentration was always 500 nM. Time in min:s.
(G) Organizational phase spaces summarizing the different experimental outcomes of KIF11-mediated microtubule network organization as a function of KIF11 and tubulin concentrations (left) and as a function of CAMSAP3-C and tubulin concentrations (right). Both plots pool the outcomes of the same 60 self-organization reactions. Temperature was 33°C.