Depletion of Fer increased the dynamics of synaptic vesicle puncta movement. (A) Live cell imaging of 14 DIV cultured hippocampal neurons that were transfected at 10 DIV with synaptophysin-GFP together with control or Fer shRNA vector. Images were captured every 60 s, and frames at 240-s intervals from one representative control and two Fer shRNA–transfected neurons are presented. (B) Kymographs represent the movement of synaptophysin-GFP along the axons. (C) A histogram represents the distribution of the speed of individual synaptophysin-GFP puncta. (D) Mean speeds of synaptophysin-GFP puncta in axons of control and shRNA-transfected neurons. n = 61 for each condition. ***, P < 0.0001. Error bars represent SEM. Bar, 5 μm.