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Fig. 1. Cell polarization in chick embryo fibroblasts. (A) Still images from a phase-contrast movie of fibroblast polarization. Time is hours:minutes:seconds. Lat A was added at –1:07:03 for just over 1 hour: within minutes in the drug, the cell rounded up (–1:00:44 to 0:00:00) and remains rounded and retracted in Lat A (0:00:00). Lat A was washed out at 0:00:00. The cell re-spreads (0:00:00 to 0:15:00), then a small stable localized retraction of the cell edge occurs (by 0:35:00, black arrow). The retracted area elaborates to form the cell rear (0:58:30, black arrows) then net protrusion and cell migration occur in the direction of the white arrow (1:06:30; net protrusion occurring for 6 minutes by this time – see E). Further migration then occurs (1:27:02). Red and green dots (in 0:15:00 to 1:27:02) mark the position of the fully re-spread cell (at 0:15:00) at the presumptive location of where the rear initiates and front forms, respectively, determined from the movie. (B) Trace of the cell in A showing direction of polarity (arrow) before and after cell repolarization. (C) Quantification of the cell population showing direction of polarity for individual cells before and after repolarization (n=15 live-cell movies). (D) Quantification of cell polarization with time after washout (% total of 15 live cells). (E) Line graph showing cell retraction to form the cell rear and protrusion at the cell front of the live cell in A measured from the red (see above) and green (see above) dot, respectively, during cell polarization (the process of re-spreading is not shown on the graph). Note that net protrusion lags net retraction. (F) Histogram of time (to nearest minute) that individual cells in the population initiate cell polarization, as identified by initial, stable, localized cell-edge retraction (n=15 live-cell movies). (G) Delay in individual cells between initial cell-edge retraction and net protrusion at the cell front (n=15 live-cell movies). Bar, 10 µm.
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