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Blood, Vol. 106, Issue 13, 4076-4085, December 15, 2005
Differential roles of microtubule assembly and sliding in proplatelet formation by megakaryocytes
Blood Patel et al.
106: 4076
Supplemental materials for: Patel et al
Files in this Data Supplement:
- Figure S1. Activation of elongation in a Triton X-100 permeabilized proplatelet by ATP (JPG, 59.4 KB)
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Changes in proplatelet length after the addition of ATP were monitored by microscopy. (A) A proplatelet viewed with DIC optics after detergent permeabilization. Two permeabilized proplatelets can be observed extending towards the right. (B) Time-lapse sequence of the lower proplatelet (arrow in A) after the addition of 1 mM ATP. ATP causes the microtubules to extrude from the end (arrows) of the permeabilized. The rate of elongation in this example is ~0.7 µm/min. Scale bar, 5 µm. See Supplementary Video 8.
- Video 1. Differential-interference-contrast microscopy time-lapse sequence showing a proplatelet elongating from a mouse megakaryocyte (MOV, 3 MB)
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The proplatelet narrows as it elongates. Images were taken every min.
- Video 2. Fluorescence time-lapse sequence of a proplatelet-containing megakaryocyte expressing EB3-GFP (MOV, 4.82 MB)
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EB3-GFP comets distribute throughout the entire length of the proplatelet and are abundant in the cell body. Both anterograde and retrograde movements of comets are apparent. Images are every 5 seconds.
- Video 3. Fluorescence time-lapse sequence of a proplatelet expressing EB3-GFP (MOV, 4.66 MB)
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EB3-GFP comets are observed moving through and around the proplatelet tips and swellings. Images are every 5 seconds.
- Video 4. Fluorescence time-lapse sequence of a proplatelet-containing megakaryocyte expressing EB3-GFP (MOV, 5.9 MB)
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EB3-GFP comets move rapidly around the proplatelet tips and throughout the cell body. Images are every 5 seconds.
- Video 5. Fluorescence time-lapse sequence of a pre-proplatelet megakaryocyte expressing EB3-GFP (MOV, 3.73 MB)
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Microtubule assembly in pre-proplatelet megakarytocytes is predominantly observed from the centrosome. EB3-GFP comets are also observed in the megakaryocyte cortex where microtubules grow in parallel with the plasma membrane. Images are every 2 seconds.
- Video 6. DIC time-lapse microscopy sequence of a proplatelet elongating in the presence of 500 nM nocodazole (MOV, 3.25 MB)
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Images are every min.
- Video 7. DIC time-lapse sequence showing a representative permeabilized proplatelet elongating following the addition of ATP (MOV, 5.37 MB)
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Images are every second.
- Video 8. DIC time-lapse sequence showing a representative permeabilized proplatelet elongating following the addition of ATP (MOV, 1.62 MB)
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Images are every second.
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