CAMSAP3-C Concentration Influences Microtubule Self-Organization by Affecting Microtubule Growth Speed and Density
(A) Confocal fluorescence microscopy images showing a time course of KIF11-mGFP-dependent organization of CF640R-labeled microtubules at different mCherry-CAMSAP3-C concentrations. Tubulin and KIF11 are present at 30 μM and 27 nM, respectively. Time in min:s.
(B) Box-and-whiskers plot depicting microtubule plus-end growth speeds at different CAMSAP3-C concentrations. 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 plus-end growth episodes measured at different mGFP-CAMSAP3-C concentrations: 0 nM, 61; 250 nM, 148; 500 nM, 72; 1,000 nM, 186. The same source data has been used for the 0 nM and 250 nM condition as for Figure 1D. The shaded area indicates the typical range of KIF11-dependent microtubule transport speeds as estimated from microtubule gliding assays in the same buffer (Figure S2E).
(C) TIRF microscopy images of 25 nM Alexa546-EB3 tracking growing microtubule ends showing enhanced microtubule formation at increasing mGFP-CAMSAP3-C concentrations at 30 μM tubulin (imaged at 2 min 20 s after initiating microtubule nucleation). Temperature was 33°C.