download.mov (8,5 MB)
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movie 1. Animated
3-D reconstruction from a stack of fluorescence images of human
CK13-EGFP chimera HK13-1 in stably transfected AK13-1 cells during
interphase. Note the typical CKF organization with thick perinuclear
cables extending toward the cell periphery to desmosomal contact sites
and the relative scarcity of filament bundles in cortical domains of
the free cell edge. |
download.mov (4,4 MB)
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movie 2. Animated
3-D reconstruction from a stack of fluorescence images of human
CK13-EGFP chimera HK13-1 in stably transfected AK13-1 cells during
metaphase. No filaments are seen but instead strong, diffuse
fluorescence is detectable throughout the cytoplasm, and aggregated
material is seen in the form of variably shaped granules and short rods
predominantly in the cortical region. |
download.mov (10 MB)
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movie 3. Fluorescence
microscopy showing stages of CKF formation at the end of mitosis in
A-431 clone AK13-1 stably expressing human CK13-EGFP chimera HK13-1. At
time point 0 min the cell has entered cytokinesis and formation of the
cleavage furrow is in progress. Granules and rodlets are seen in the
cytoplasm and are enriched in the cell cortex at that time. Some
cytoplasmic rodlets further elongate (2.5 min) but disappear later
(22.5 min). Rebuilding of a typical CKF network originates from the
cell periphery and proceeds toward the cell center. Note that diffuse
cytoplasmic fluorescence decreases during filament formation.
Simultaneously, cells flatten and re-establish extensive contacts with
neighboring cells. |
download.mov (6 MB)
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movie 4. Using
confocal scanning laser microscopy images were obtained from human
vulvar carcinoma-derived A-431 subclone AK13-1 stably expressing
fluorescent CKFs containing human CK13-EGFP chimera HK13-1. The movie
shows five focal planes from bottom to top. At the beginning of the
recording (early prophase) partial filament breakdown has already taken
place in central cytoplasmic regions and increased diffuse cytoplasmic
staining can be seen. At that time point the nucleus is still intact.
Filament disassembly continues from the center of the cell towards the
periphery, where the originally fine network concentrates in more solid
structures and disintegrates into granules. Note, that at the end of
the sequence CKFs are completely absent. Instead, significant diffuse
cytoplasmic staining and granules, most of which are located in the
cell periphery, are detected in the rounded cells. At some intermediate
time points rodlets arise in the cytoplasm (e.g., level 2, 7.5 min) but
disappear later. |
download.mov (3,3 MB)
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movie 5. Time-lapse
fluorescence microscopy of dividing AK13-1 cell depicting the
distribution of human CK13-EGFP chimera HK13-1 in the absence of drugs.
The movie taken as control shows only one of several confocal image
planes. |
download.mov (1,8 MB)
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movie 6. Time-lapse
fluorescence microscopy of dividing AK13-1 cells depicting the
distribution of human CK13-EGFP chimera HK13-1 in the presence of 0.05%
[w/v] sodium azide and 50 mM deoxyglucose. The movie shows only one of
several confocal image planes. Note the differences to the control
movie 5. |
download.mov (4,8 MB)
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movie 7. Time-lapse
fluorescence microscopy of dividing AK13-1 cells depicting the
distribution of human CK13-EGFP chimera HK13-1 in the presence of
cytochalasin D (2 µM) to disrupt actin filaments. The movie shows only
one of several confocal image planes. Note differences to control movie
5. |
download.mov (3,5 MB)
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movie 8. Time-lapse
fluorescence microscopy of dividing AK13-1 cells depicting the
distribution of human CK13-EGFP chimera HK13-1 in the presence of
nocodazole (1 µM) to disrupt microtubules.The movie shows only one of
several confocal image planes. Note the differences to the control
movie 5. |
download.mov (2,5 MB)
|
movie 9. Time-lapse
fluorescence microscopy of dividing AK13-1 cells depicting the
distribution of human CK13-EGFP chimera HK13-1 in the presence of the
tyrosine phosphatase inhibitor pervandate (20 mM). The movie shows only
one of several confocal image planes. Note the differences to the
control movie 5. |
