AnkB interaction with p62 regulates the motility of multiple axonal cargoes and growth of axons. (A) For each genotype the top represents the distribution of synaptophysin-YFP vesicles and 220-kD WT or DD1320AA AnkB-mCherry in DIV7 axons. Arrowheads indicate large axonal aggregates. (middle) Kymograph analysis of the motion of all particles. (bottom) Corresponding color-coded trajectories for a subset of YFP-positive particles show static vesicles (blue) and anterograde (green)- and retrograde (red)-moving vesicles. (B and C) Axonal velocity (B) and run length (C) for the indicated cargoes. The data shown are from a single representative experiment out of five repeats. For the experiment shown, data were computed from n = 10 axons from five independent neuronal preparations (for synaptophysin-YFP, anterograde [n = 175] and retrograde [n = 150] YFP-particles; for other organelles, see supplemental figure legends). (D) Representative images of DIV8 hippocampal neurons expressing dsRed alone or in combination with WT or DD1320AA AnkB-GFP (arrowheads indicate axons). (E) Quantification of axonal length at DIV8 (n = 30 cells). The data shown are from a single representative experiment out of three repeats. Data represent means ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA with Tukey’s post-test.