Myosin-Vb functions as a dynamic tether for peripheral endocytic compartments during transferrin trafficking

Additional file 1:

Low-level expression of full-length eGFP-myosin-Vb (green) shows rare, dynamic colocalization (circles) of myosin-Vb and transferrin (red); same cell as shown in Figure 1D–F. Frame acquisition rate, 0.5/sec; elapsed seconds are displayed at lower right.

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Additional file 2:

Overexpression of full-length eGFP-myosin-Vb (green) in the presence of transferrin (red; added at the time of transfection) produces enlarged, less-motile peripheral endosomes decorated with myosin-Vb and containing transferrin at 24 h post transfection; same cells as shown in Figure 2D–F. Frame acquisition rate, 0.5/sec; elapsed seconds are displayed at lower right.

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Additional file 3:

Overexpression of full-length eGFP-myosin-Vb produces enlarged, less-motile peripheral endosomes decorated with myosin-Vb; same field as shown in Figure 2J–L. Frame acquisition rate, 0.5/sec; frame display rate, 3/sec.

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Additional file 4:

Overexpression of full-length eGFP-myosin-Vb (not shown) prevents entry of Alexa 546-labeled transferrin (shown) into perinuclear compartments; same field as Additional file 3. Frame acquisition rate, 0.5/sec; frame display rate, 3/sec.

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Additional file 5:

Overlay of Additional file 3 (myosin-Vb, green) and Additional file 4 (transferrin, red).

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Additional file 6:

Rab11a (red) colocalizes with eGFP-myosin-Vb (green) at high myosin-Vb expression levels. Frame acquisition rate, 0.5/sec; frame display rate, 6/sec.

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Additional file 7:

Rab4 (red) does not colocalize with eGFP-myosin-Vb (green) at high myosin-Vb expression levels. Frame acquisition rate, 0.5/sec; frame display rate, 6/sec.

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Additional file 8:

Rab5 (red) does not colocalize with eGFP-myosin-Vb (green) at high myosin-Vb expression levels. Frame acquisition rate, 0.5/sec; frame display rate, 6/sec.

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Additional file 9:

Chemical-genetic inhibition of sensitized mutant (Y119G) eGFP-myosin-Vb by PE-ADP microinjection does not prevent movement of transferrin-positive particles. Cells were loaded with fluorescent transferrin (red) 30 min before myosin-Vb was inhibited in the center cell by PE-ADP. Frame acquisition rate, 1/sec; frame display rate, 3/sec.

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Additional file 10:

Chemical-genetic inhibition of sensitized mutant (Y119G) eGFP-myosin-Vb by PE-ADP microinjection (cell on left) halts movement of all myosin-Vb-decorated particles, including those being transported via microtubules; same field as Figure 3A. Uninjected control cell is on the right. Frame acquisition rate, 1/sec; frame display rate, 10/sec.

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Additional file 11:

Same conditions as Additional file 10 without a control uninjected cell. Frame acquisition rate, 1/sec; frame display rate, 10/sec.

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Additional file 12:

Negative control cell expressing wild-type eGFP-myosin-Vb; PE-ADP injection (immediately before imaging) does not halt movement of myosin-Vb-decorated particles. Frame acquisition rate, 1/sec; frame display rate, 10/sec.

Format: MOV Size: 7.1MB Download file

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