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Supplementary Movie Legends
Page 1
Kamei et al., MS# 2006-01-00604A
SUPPLEMENTAL MOVIE LEGENDS
Supplemental Movie 1
This movie shows transmitted light time-lapse images of several human EC in a 3D
collagen gel forming numerous intracellular vacuoles, some of which combine to form
highly enlarged spaces within and between the cells. The time-lapse covers a period of
about 11 hours, with approximately 60 minutes elapsed time per second of movie
(QuickTime; 2.6 MB). Selected frames from this movie are shown in Figure 2a.
Supplemental Movie 2
This movie shows transmitted light time-lapse images of a single human EC in a 3D
collagen gel forming a greatly enlarged intracellular ”lumenal” space encompassing the
vast majority of the inside of the cell. The time-lapse covers a period of about 24 hours,
with approximately 100 minutes elapsed time per second of movie (QuickTime; 2.7 MB).
Selected frames from this movie are shown in Figure 2b.
Supplemental Movie 3
This movie shows two-photon time-lapse images of a ventral intersegmental endothelial
cell in the trunk of a Tg(fli1:EGFP-cdc42wt)y48 transgenic zebrafish. Vacuoles rapidly
appear, rearrange, and disappear within the cell. The time-lapse covers a period of about
4-1/2 hours, with approximately 15 minutes elapsed time per second of movie
(QuickTime; 4.3 MB). Each movie frame is a reconstruction from a Z-series of 15-18
planes (with 2 µm between planes) collected at that time point. Selected frames from this
movie are shown in Figure 2c.
Supplementary Movie Legends
Page 2
Kamei et al., MS# 2006-01-00604A
Supplemental Movie 4
This movie shows two-photon time-lapse images of a ventral intersegmental endothelial
cell in the trunk of a Tg(fli1:EGFP-cdc42wt)y48 transgenic zebrafish. A single vacuole
within the cell enlarges and expands to become an enlarged lumenal space, eventually
forming a ventral connection to the lumen of the dorsal aorta below. The time-lapse
covers a period of about 3 hours, with approximately 30 minutes elapsed time per second
of movie (QuickTime; 4.2 MB). Each movie frame is a reconstruction from a Z-series of
15-18 planes (with 2 µm between planes) collected at that time point. Selected frames
from this movie are shown in Figure 2d.
Supplemental Movie 5
This movie shows two-photon time-lapse images of the ventral portion of a developing
intersegmental vessel in the trunk of a Tg(fli1:EGFP-cdc42wt)y48 transgenic zebrafish.
Vacuoles appear within the cell and merge and coalesce to form a gradually enlarging
lumenal space. The time-lapse covers a period of about 11-1/2 hours, with approximately
30 minutes elapsed time per second of movie (QuickTime; 8.9 MB). Each movie frame
is a reconstruction from a Z-series of 15-20 planes (with 2 µm between planes) collected
at that time point. Selected frames from this movie are shown in Figure 2e.
Supplemental Movie 6
This movie shows two-photon time-lapse images of the ventral portion of a developing
intersegmental vessel in the trunk of a Tg(fli1:EGFP-cdc42wt)y48 transgenic zebrafish.
Vacuoles appear within the cell and merge and coalesce to form a gradually enlarging
lumenal space. The time-lapse covers a period of about 11.5 hours, with approximately
Supplementary Movie Legends
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Kamei et al., MS# 2006-01-00604A
30 minutes elapsed time per second of movie (QuickTime; 8.9 MB). Each movie frame
is a reconstruction from a Z-series of 17-23 planes (with 2 µm between planes) collected
at that time point.
Supplemental Movie 7
This movie shows transmitted light time-lapse images of two initially separate human EC
in a 3D collagen gel. The two cells form intracellular vacuoles that combine to form a
single large large lumenal space bounded by both cells. The time-lapse covers a period
of about 19 hours, with approximately 100 minutes elapsed time per second of movie
(QuickTime; 1.7 MB). Selected frames from this movie are shown in Figure 3a.
Supplemental Movie 8
This movie shows transmitted light time-lapse images of a group of human EC in a 3D
collagen gel. The cells sequentially incorporate into a multicellular group of cells
bounding a single enlarging lumenal space. The accompanying movie 9 shows that the
formation of this structure is accomplished without mixing of the cytoplasmic contents of
the aggregating cells. The time-lapse covers a period of about 40 hours, with
approximately 120 minutes elapsed time per second of movie (QuickTime; 9.4 MB).
Each movie frame is a reconstruction from a Z-series of 4 planes (with 5 µm between
planes) collected at that time point. Selected frames from this movie are shown in Figure
3c (bottom).
Supplemental Movie 9
This movie shows composite red/green epifluorescence time-lapse images of a mixed
group of differentially labeled human EC in a 3D collagen gel. The cells sequentially
Supplementary Movie Legends
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Kamei et al., MS# 2006-01-00604A
incorporate into a multicellular group of cells bounding a single enlarging lumenal space.
The accompanying movie 8 shows the corresponding transmitted light images of the
same field of cells. Note that the frames in movie 9 only correspond to every sixth
frame/time point in movie 8. The time-lapse covers a period of about 40 hours, with
approximately 120 minutes elapsed time per second of movie (QuickTime; 7.9 MB).
Each movie frame is a reconstruction from a Z-series of 4 planes (with 5 µm between
planes) collected at that time point. Selected frames from this movie are shown in Figure
3c (top).
Supplemental Movie 10
This movie shows a composite red/green time-lapse two-photon images of the ventral
portion of a developing intersegmental vessel in the trunk of a Tg(fli1:EGFP-cdc42wt)y48
transgenic zebrafish injected intravascularly with 605 nm quantum dots. 960 nm twophoton excitation permits simultaneous activation of EGFP (green) and quantum dot
(red) emission, which are then collected by separate PMT detectors. The movie shows
successive labeling of preformed vacuolar compartments in the ventral-most endothelial
cell and then the next more distal endothelial cell via transfer of quantum dots that are
initially present only in the dorsal aorta. The time-lapse covers a period of about 2 hours,
with approximately 10 minutes elapsed time per second of movie (QuickTime; 7.5 MB).
Each movie frame is a reconstruction from a Z-series of 15 planes (with 2 µm between
planes) collected at that time point.