Cross-linking decreases MT comet velocities throughout the cell, limits MT entry into the peripheral zone, and increases the percent of comets that are swept along the peripheral zone boundary. (A) Schematic of MT comet trajectories categorized in S2 cells (left) and change in mean comet velocity in cross-linked compared with non–cross-linked cells (right, based on mean velocities presented in B). (B) EGFP comet velocities from control cells (cotransfected with SKIP-LZ-iLID^Lit + CH-CH, open circles) and constitutively cross-linked cells (cotransfected with SKIP-LZ-iLID^Lit + CH-CH-SspB, filled circles), grouped based on comet trajectory as delineated in A. Multiple individual EGFP-labeled comet velocities were determined and compiled across multiple cells. Numbers in parentheses indicate (number of independent experiments, total number of cells analyzed, total number of tracks annotated). Mean comet velocities are reported below the dotted line, and the relative percentages of comets in each category are shown at the top of the plot (bold percentages: >2-fold difference between cross-linked and non–cross-linked cells). Line represents the mean, and error bars indicate SD. P-values were determined by two-way unpaired Student’s t test. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001.