Knockdown of NudE and LIS1 resembles acute inhibitions. (A) Lysos/LEs in rat hippocampal neurons moved mostly in the retrograde direction in wild-type and scrambled RNAi controls, but an increase in bidirectionally moving lysosomes and severe immobility of larger lysosomal particles (≥1 µm in diameter) were apparent in NudE RNAi and LIS1 RNAi cells, respectively. (A–C) Consistent with the NudE/L antibody–injected case, knockdown of NudE caused an increase in bidirectionally moving lysos/LEs (A and B), whereas knockdown of LIS1 protein more severely interfered with mobility of larger particles than smaller ones, which is consistent with the LIS1 antibody–injected case (A–C). Stationary, anterograde/retrograde, and bidirectional particles were defined as particles that displaced |ΔX| < 1 µm, >5 µm in one direction, and >3 µm in both directions, respectively. The error bars represent SD (*, P < 0.001). The number of particles analyzed are as follows: control (small), n = 60; control (large), n = 55; LIS1 RNAi (small), n = 40; LIS1 RNAi (large), n = 66; NudE RNAi (small), n = 46; and NudE RNAi (large), n = 38 (using 10 axons per condition).