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. Author manuscript; available in PMC: 2012 May 12.
Published in final edited form as: Neuron. 2011 May 12;70(3):441–454. doi: 10.1016/j.neuron.2011.03.022

Figure 3. Biased transport of cytosolic cargoes is distinct from untagged PAGFP and is motor-dependent.

Figure 3

(A) To determine the effects of NEM on generic motor-driven transport, neurons were transfected with APP:YFP and transport of APP vesicles was determined before and after adding 0.5μM NEM. Each panel on left is created by overlapping three consecutive time-lapse frames that were pseudo-colored red/blue/green respectively; before (top frame) and at incremental time-points after adding NEM. Corresponding kymographs from the movies are shown on the right. Note that the robust bidirectional vesicular transport was gradually inhibited with NEM-treatment. (B) Neurons transfected with PAGFP:synapsin or PAGFP:CamKIIa were incubated with 0.5mM NEM for 10 minutes and photoactivation experiments were performed as described in text. As shown in the representative kymographs (top), NEM treatment greatly inhibited the overall mobility of the photoactivated pool, leading to stalled fluorescent particles within axons and essentially eliminated the anterograde bias, as shown in the graphs below (mean ± SEM). Scale bars: lower right, elapsed time in seconds: right/left of kymograph. (C) A similar inhibition of the anterograde bias of synapsin was seen upon treatment of neurons with the microtubule-depolymerizing drug Nocodazole (mean ± SEM, p<0.0001, paired t-test).