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Fig. 6. Dynamics of fluorescent transferrin and LysoTracker labelled organelles in cells depleted of lamin A/C, p150^Glued, CLIP-170 or EB1. (A) Live HeLa cells depleted of lamin A/C, p150^Glued, CLIP-170 or EB1 were labelled with Alexa Fluor 568-transferrin for 10 minutes and subsequently imaged at three frames per second. The image shows enlarged regions of cells with tracks of moving objects overlayed onto them. Bar, 5 µm. See also supplementary material Movie 1. (B) Tracking of Alexa Fluor 568-transferrin-positive structures reveals that the speed of movement is unchanged by depletion of either Lamin A/C, p150^Glued, CLIP-170 or EB1. Objects from at least 20 cells from two independent experiments were analyzed; histograms show the average of this data, the points plotted show the spread of velocities from 25 randomly selected objects of the >500 objects tracked. (C) Live HeLa cells depleted of lamin A/C, p150^Glued, CLIP-170 or EB1 were labelled for 5 minutes with LysoTracker and subsequently imaged at two frames per second for 1 minute. The image shows cells with tracks of moving objects overlayed onto them. Bar, 10 µm. See also supplementary material Movie 2. (D) Tracking of LysoTracker-positive structures reveals that the speed of movement is unchanged by depletion of either Lamin A/C, p150^Glued, CLIP-170 or EB1. Objects from at least ten cells for each depletion from two independent experiments were analyzed; histograms show the average of this data, the points plotted show the spread of velocities from 25 randomly selected objects of the >200 objects tracked for each depletion experiment. Stars in A and C indicate starting positions of the objects being tracked.

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