Perturbation of MT-based motility abrogates directed VIF transport. (A) 15-s vimentin-SunTag trajectories in an untreated control cell, a nocodazole-treated cell (10 µM), a KIF5B KO cell, a dynapyrazole A–treated cell (5 µM), and a p50-overexpressing cell. Motile trajectories with maximum displacements >1 µm are shown in red; all other trajectories are shown in gray. The percentage of motile tracks in each representative example is labeled in red. Scale bars = 10 µm. (B) Percentage of all 15-s vimentin-SunTag trajectories in each condition classified by maximum displacement into stationary (≤0.5 µm, dark gray), intermediate (0.5–1.0 µm, light gray), or motile (>1 µm, red) groups. Total tracks analyzed per condition are indicated on the left. (C) Percentage of motile vimentin-SunTag trajectories binned per cell for each condition. (D) Origin-aligned motile trajectories (>1 µm) for each condition. Trajectories are randomly colorized for visibility. Analysis was conducted on a total of 20 individual cells for each condition, collected from three independent experiments. KO, knockout.