Detailed view of time-lapse microscopy of Spastin-mediated microtubule severing. NIH 3T3 cells stably transfected with GFP-tubulin were permeabilized as described in Materials and methods and 80 nM WT Spastin was added. After selecting a field to image, 0.25 mM ATP was added at time 0. The elapsed time is stamped on each frame and the total time is ~9 min. The video shows a detail view of a region (same region as Fig. 4 e) of the cell shown in Video 1. Frames from this movie are also shown in Fig. 4. Images were acquired at no more than 15-s intervals. Elapsed times are stamped in each frame. The frame display rate is 15 frames/s.
