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Fig. 8. GFP-actin dynamics during wound healing of MTD1-A cells. (A) Confluent monolayers of MTD1-A cells stably expressing GFP-actin were scratched with a needle (18 gauge) after a brief incubation in PBS. Time-lapse images of GFP-actin (single confocal sections) were acquired by confocal microscopy at an appropriate time after wounding (see supplementary material Movie 6). The images shown are still-frames from the time-lapse data at the indicated times. Note that rapid actin polymerization occurs at the tips of the leading edges immediately after the initial cell-cell contacts (arrowheads), and the resultant radial actin fibers tag the circumferential actin cables when they come into contact (00:35). (B) The images in A were subjected to kymograph analysis to monitor the changes in the distance between the two circumferential actin cables in contacting cells along the broken line. Arrow, indicates the time point when radial actin fibers reached the intracellular circumferential actin cables. (C) GFP-actin dynamics at the late stage of wound healing. Note that the radial actin fibers (traced by magenta lines in the bottom panels) are aligned and fused with the corresponding circumferential actin cables (illustrated by white lines in the bottom panels) (see supplementary material Movie 7). Scale bars: 10 µm.
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