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Supplemetal figures
Supplemental Figure 1
Effects of nocodazole and latrunculin B on the microtubule and actin
cytoskeleton. (a) Immature oocytes were incubated for 30 min in 3.3 μM
nocodazole and processed for immunofluorescence. (b) Control cells and
oocytes treated with 250 nM latrunculin B were matured and fixed 5 min after
NEBD. Scale bars: 20 μm. (Adobe Acrobat PDF, 700 kB)
Supplemental Figure 2
Low doses of LAB also strongly delay chromosome movement. (a) As
Figure 2c-e, Z-projections of selected time points from a 4D dataset (25×3µm
every 40s). 120 nM Latrunculin B was added 10 min before NEBD. Time:
mm:ss, 0:00: NEBD; scale bar 20 μm. Full movie and isosurface reconstruction:
MovieS2. (b) 3D chromosome tracks. (c) Distances of chromosomes from the
animal pole calculated from (b). (Adobe Acrobat PDF, 140 kB)
Supplemental Figure 3
DNA coated beads nucleate actin patches similar to chromosomes. (a)
DNA-coated beads (green) and Alexa 488 actin (red) were injected into the
nucleus and cytoplasm of an oocyte, respectively. (b) MBP (maltose binding
protein) blocked beads. DIC images clearly indicate the location of the only
weakly fluorescent beads. (a,b) Each frame is a projection of three optical
sections. Time mm:ss, 0:00: NEBD; scale bar 20 μm. (Adobe Acrobat PDF, 3.6
MB)
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Supplemental videos
Video S1
Microtubules are too short to capture chromosomes in meiosis I of
starfish oocytes. Microtubules: red; chromosomes: green. Each frame is a
projection of 20 optical sections. (QuickTime, 3.2 MB)
Video S2
Chromosome congression in an untreated oocyte. Chromosomes were
labeled by expressing H2B-diHcRed and imaged in 4D over maturation. Left:
projections of the raw data; right: isosurface reconstruction and tracks of
chromosomes. (QuickTime, 3.1 MB)
Video S3
Chromosome congression in an oocyte preincubated for 1h and matured
in 3.3 μM nocodazole. Chromosomes were labeled by expressing H2BdiHcRed and imaged in 4D over maturation. Left: projections of the raw data;
right: isosurface reconstruction and tracks of chromosomes. (QuickTime, 2.2
MB)
Video S4
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Chromosome congression in an oocyte treated with 2 μM LAB 10 minutes
before NEBD. Chromosomes were labeled by expressing H2B-diHcRed and
imaged in 4D over maturation. Left: projections of the raw data; right: isosurface
reconstruction and tracks of chromosomes. (QuickTime, 3.6 MB)
Video S5
Chromosome congression in an oocyte treated with 3.3 µM nocodazole
and 250 nM LAB at the time of hormone addition. Chromosomes were
labeled by injecting recombinant Alexa 568 labeled Ran and imaged in 4D over
maturation. Left: projections of the raw data; right: isosurface reconstruction and
tracks of chromosomes. (QuickTime, 4.1 MB)
Video S6
A contractile actin meshwork moves chromosomes to the animal pole in
starfish oocytes. F-actin (GFP-ABD): gray, chromosomes (Alexa 568 Ran):
red. Each frame is a projection of three optical sections. Note the capture of
chromosomes after the depolymerization of the surrounding actin patches.
(QuickTime, 5.3 MB)
Video S7
Details of the actin mesh. Single optical sections from a GFP-ABD expressing
oocyte. Note the contraction of the actin network towards the animal pole.
(QuickTime, 2.4 MB)
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Video S8
Phalloidin injection delays the collapse of the actin mesh. Single optical
sections from a H2B-GFP expressing oocyte injected with TMR-phalloidin ~3
min after NEBD. (QuickTime, 2.0 MB)
Video S9
Chromosome congression in an oocyte treated with 120 nM LAB 10
minutes before NEBD. Chromosomes were labeled by expressing H2BdiHcRed and imaged in 4D over maturation. Left: projections of the raw data;
right: isosurface reconstruction and tracks of chromosomes. (QuickTime, 1.7
MB)
Video S10
DNA coated beads nucleate actin patches similar to chromosomes. Green:
DNA attached to beads; red: Alexa 488 actin. Each frame is a projection of
three optical sections. (QuickTime, 1.2 MB)
Video S11
Uncoated beads do not nucleate actin. 1 μm beads blocked with maltose
binding protein injected into oocyte nuclei. Green: autofluorescence of beads;
red: Alexa 488 actin. Each frame is a projection of three optical sections. As
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autofluorescence of beads is weak, DIC images are shown to the right for
clarity. (QuickTime, 2.7 MB)