Fig. 2
Microtubule (MT) wall-to-end transition by CENP-E paired with Ndc80 complex. a Schematics of the MT wall-to-end transition assay and a representative imaging field with GMPCPP-stabilized MTs (red) and coverslip-immobilized beads coated with green fluorescent protein (GFP)-labeled Ndc80 protein (green). Bar, 3 µm. b Selected images showing motions of MTs on immobilized beads coated with the indicated proteins. Numbers are time (min) from the start of observation. Arrows show direction of MT gliding. Bar, 3 µm. c Quantifications for the wall-to-end transition assay using a mixture of CENP-E motors and different Ndc80 proteins. Columns are means ± SD for results from N independent trials, which are shown with gray dots. For CENP-E only, CENP-E paired with either Ndc80 Broccoli, Ndc80 Bonsai, Δ1–80 Ndc80 Bonsai, or K166D Ndc80 Bonsai, N = 4, 7, 2, 3, and 2, respectively. Total number of examined MTs in all trials is indicated below each column. Source data are provided as a Source Data file. A successful MT end-retention event was counted if the trailing MT end was coupled to the bead for longer than 4 s. All Ndc80 proteins showed high percent of end-retention events but their durations were dramatically different. d Kaplan–Meier survival plot for MT end-retention time based on N independent trials examining end-attachment for n MTs: CENP-E motor: N = 4, n = 23; Ndc80+CENP-E, N = 7, n = 111
