JCS017640 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Organization of adhesion complexes during cell polarization. Cell polarization was initiated in primary chick fibroblasts expressing GFP-actin; then, at various times during polarization cells were fixed and stained with anti-talin antibody and overlay images of the two protein patterns were obtained.

• Movie 1 -

  Movie 1. Phase-contrast movie of cell polarization for an individual fibroblast. Time (in hours:minutes:seconds) is given before (−) and after Lat A washout. At the start of the movie the fibroblast is untreated and polarized and migrating. Lat A is added at −1:07:03. This induces cell rounding and abolishes cell polarity. At 0:00:00 Lat A is washed off to initiate cell polarization. Still images from the movie are shown in Fig.1.

• Movie 2 -

  Movie 2. A fibroblast expressing GFP-actin imaged live during cell polarization initiated by Lat A washout. Time given is the actual time of movie acquisition (hours:minutes:seconds). At 13:43:14 Lat A had been washed out for 2 minutes (00:02:00). Actin in the cell body polymerizes and then reorganizes to form oriented actin filament bundles. Formation of oriented actin filament bundles occurs prior to cell polarization and migration. Still images from the movie are shown in Fig. 3.