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Files in this Data Supplement:
Fig. S1. Confocal z slices of the cell tail rupture event from Fig. 1C′. Individual confocal z slices (at 1 µm steps) for the time points indicated in Fig. 1C′, demonstrating that the cellular particle (red arrow) that remains behind near the apical surface was originally part of the mem-EGFP labeled cell (red dot) before the cell tail was retracted (red arrowhead). Scale bar: 10 µm. nt, neural tube; nc, neural crest; ap, apical.
Fig. S2. Confocal z slices of the cell tail rupture from Fig. 4C′. Individual confocal z slices (at 1 µm steps) from the time points indicated in Fig. 4C′, showing that the apical GFP-α-catenin signal (white arrowhead) that remains near the apical surface was originally part of the GFP-α-catenin labeled cell (dot) before it pulled away from the lumen (white arrow) to become a neural crest cell. The red arrowhead marks an apical GFP-α-catenin signal that is close to, but not associated with, the neural crest cell (dot) GFP-α-catenin signal (white arrowhead). Scale bar: 10 µm.
Fig. S3. Analysis of the apical mitotic cleavage plane in dorsal neural tube cells. (A-C) Scoring the angle of dorsal neural tube cell division from time-lapse imaging. The cleavage plane of apical cell division (red dashed line) was compared to the plane of the neural epithelium (green dashed line) of individual neural tube cells and categorized as being either 60-90° perpendicular to the plane of the epithelium (A), 30-60° (B) or 0-30° (C) parallel to the plane of the epithelium. The green dashed line in the boxes on the left shows the apical surface (ap). Scale bar: 10 µm. (D) Bar graphs showing the time spent in mitosis for neural tube cells that either underwent an EMT (gray bars) or remained within the neural tube (white bars). Error bars indicate s.e.m.
Movie 1. Time-lapse imaging with mem-EGFP; corresponds to Fig. 1B′. Complete detachment: neural tube cell (red dot) detaches from the apical domain of the neural tube and exits the neural tube without leaving any detectable debris behind at the apical surface of the neural tube. Here, as in all movies; ap, apical; nt, neural tube; nc, neural crest. The time points are all 7 minutes apart, dotted lines indicate the boundaries of the neural tube and dorsal is up. Scale bars: 10 µm. In QuickTime Player, frame by frame advance can be controlled with the left and right arrow keys.
Movie 2. Time-lapse imaging with mem-EGFP; corresponds to Fig. 1C′. Ruptured cell tail: neural tube cell (red dot) retracts its cell tail (red arrowhead) from the apical domain of the neural tube and a mem-EGFP labeled structure (red arrow) is left behind within the neural tube, indicating that the cell tail was ruptured as it was withdrawn. A more detailed confocal analysis of the cell tail rupture is shown in Fig. S1.
Movie 3. Time-lapse imaging with mem-EGFP; corresponds to Fig. 2A′. Non-apical mitosis: neural tube cell (red dot) undergoes nuclear migration and rounds up at the apical domain of the neural tube in mitosis. Prior to division the mitotic neural tube cell departs from the apical domain and division occurs at a non-apical location. By the end of the movie the daughter cell with the red dot has left the boundaries of the neural tube.
Movie 4. Time-lapse imaging with mem-EGFP; corresponds to Fig. 3A′. Dynamic cell-tail detachment prior to an EMT: as this neural tube cell (red dot) detaches from the apical domain of the neural tube, the cell tail is first partly withdrawn, then reinserted and then withdrawn completely.
Movie 5. Time-lapse imaging with mem-EGFP; corresponds to Fig. 3B′. Temporary neural crest cell re-attachment: a neural crest cell (red dot) sends a cellular process to the apical surface of the neural tube and then retracts it and migrates out of the field of view.
Movie 6. Time-lapse imaging with GFP-actin; corresponds to Fig. 4A′. GFP-actin reduced prior to detachment: the apical GFP-actin signal (white arrowhead) of a neural tube cell (dot) is noticeably reduced just prior to retraction of the cell tail.
Movie 7. Time-lapse imaging with GFP-α-catenin and mCherry-tubulin; corresponds to Fig. 4B′. GFP-α-catenin signal travels with the cell tail during detachment: as a neural tube cell (dot) retracts its cell tail the GFP-α-catenin signal (white arrowhead) remains at the tip of the cell tail.
Movie 8. Time-lapse imaging with GFP-α-catenin; corresponds to Fig. 4C′. GFP-α-catenin signal remains at the apical surface after detachment: as a neural tube cell (dot) retracts its cell tail (white arrow) the apical GFP-α-catenin signal (white arrowhead) remains behind at the lumen in close proximity to a different apical GFP-α-catenin signal (red arrowhead) not associated with the same cell. A more detailed confocal analysis of the cell tail rupture is shown in Fig. S2.
Movie 9. Time-lapse imaging with g-tubulin-GFP and mCherry-tubulin; corresponds to Fig. 5A′. γ-tubulin-GFP during interkinetic nuclear migration: as this cell undergoes interkinetic nuclear migration and apical mitosis, the centrosome (γ-tubulin-GFP signal) leaves the apical surface and travels basally. Minutes later the nucleus moves towards the apical domain. The nucleus meets the centrosome in a non-apical location, and then both the centrosome and the nucleus travel together to the apical surface where mitosis occurs. In this movie a periodic rotation of ±20° on the y-axis was added to show depth.
Movie 10. Time-lapse imaging with g-tubulin-GFP and mCherry-tubulin; corresponds to Fig. 5B′. γ-tubulin-GFP during the EMT: in a neural tube cell (dot) that retracts its tail (white arrowhead), the γ-tubulin-GFP signal stays at the apical-most domain of the tail.
Movie 11. Time-lapse imaging with g-tubulin-GFP; corresponds to Fig. 5C′. γ-tubulin-GFP speckles: a neural tube cell (dot) transports γ-tubulin-GFP speckles (white arrowheads) intracellularly from the nucleus to the apical domain until the moment of cell tail detachment.
Movie 12. Time-lapse imaging with mem-EGFP; corresponds to Fig. 6A′. Parallel mitosis followed by an EMT: neural tube cell (red dot) undergoes mitosis at the apical domain of the neural tube with a cleavage plane that separates the basal-most daughter cell from the apical surface. The basal-most daughter cell (red dot) then exits the neural tube to become a neural crest cell, but not before severing a midbody attachment (yellow arrowhead) and re-extending a cellular process (red arrowhead) towards the apical surface, before retracting it again.
Movie 13. Time-lapse imaging with mem-EGFP; corresponds to Fig. 6B′. Parallel mitosis with no EMT: neural tube cell (red dot) undergoes mitosis at the apical domain of the neural tube with a cleavage plane that separates the basal-most daughter cell from the apical surface. The basal-most daughter cell (red dot) is later found re-attached to the apical domain of the neural tube (red arrows), and both daughter cells remain within the neural tube.
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