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To confirm this, we also analyzed MT polarity using the direction of movement of single GFP-tagged dynein motors on the track and transport MT, in cases where the transport MT moved past the end of the track MT. In 21 of 22 cases, the transport and track MTs were anti-parallel to one another.
The transport MT is captured and aligned onto the track MT by GST-Dyn1331kDa.
Arrowhead shows the red transport MT slides with its minus-end away from the plus-end of the track MT, indicating that sliding is anti-parallel.
The transport MT pauses before reaching the end of the track MT, which was frequently observed for rat dynein-driven movement.
Loss of either Sac1 or DVAP function disrupts axonal transport, MT stability, synaptic growth and the localization of a number of postsynaptic markers.
Arrowheads follow the transport MT as it slides along the track MT. Asterisk marks where the transport MT remains attached at the end of the track MT. Right, histograms of the MT-MT sliding and surface gliding velocities driven by GST-Dyn1331kDa with Gaussian fitting.
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After addition of ATP, GST-Dyn1331kDa drobustobuslidinging of transport MTs along the immobilized track MTs.
Unbound dynein was removed by washing with BC buffer, followed by introduction of transport MTs diluted in BC buffer.
The transport MTs frequently moved until they reached the end of the track MT, where they swiveled around an end point on the track MT (asterisk Figure 2A).
In this revision, we have included an additional method of assessing polarity- direct single molecule observation of the direction of movement of individual GFP-dynein molecules on the track and transport MTs.
We have also distinguished the longer MTs and shorter transporting MTs with two different colors as the reviewers suggested.
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