Kip2 residues K294 and R296 are dispensable for Kip2 motility in vivo and in vitro. (A) Representative preanaphase cells (T1: the first frame), kymographs drawn from time-lapse series, and highlighted trajectories (red dashed lines) of Kip2-3xsfGFP (top) and Kip2-P1−-3xsfGFP (bottom) speckles moving along a metaphase aMT. Spc72-GFP was used to visualize SPBs. KIP3 was deleted to increase the aMT length. Scale bars, 2 μm. (B) Quantification of Kip2 speckle moving speed using the kymographs shown in A. More than 90 speckles analyzed per condition. (C) Representative kymographs showing random landing along GMPCPP-stabilized microtubule lattices, processive motility, and plus-end accumulation of individual Kip2-WT and Kip2-P1− motors. (D) Quantification of Kip2 motor speed using the kymographs shown in C. More than 55 speckles analyzed per condition. Statistical significance was calculated using two-tailed Student's t test; * P < 0.05; ** P < 0.01. Source data for C and D are available in Data S1.
