JCS01734 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Migration of B16F1 populations on laminin. Cells expressing vinculin-wt and -LD were plated on laminin-coated coverslips and subjected to video microscopy 10-14 hours after seeding. Selected frames from movies of representative cells, taken in fluorescence mode, were pseudocoloured red (t=0 minutes) and green (t=11.5 minutes), and merged to illustrate the locomotory activity of representative cells. Scale bars, 10 mm. Notice that the (small) adhesions of the motile vinculin-wt-expressing cells were completely translocated and reorganized after 11.5 minutes, whereas the adhesions in vinculin-LD-expressing cells were largely maintained. In addition, focal adhesions of vinculin-LD-expressing cells were larger than those of motile vinculin-wt-expressing cell (also compare Fig. 2).

• Supplemental Figure 2 -

  Fig. S2. Effects of activated Rac on B16-F1 cells. B16-F1 populations expressing GFP-vinculin variants were seeded onto fibronectin, transfected with activated Rac or L61-Rac, and fixed 12 hours after transfection. Localization of GFP-vinculin-wt or GFP-vinculin-LD, L61-Rac and F-actin are shown individually (greyscale) and in merged images. Scale bars, 10 mm. Notice the flat, spread-out morphology of B16-F1 cells expressing L61-Rac with peripheral ruffles. GFP-vinculin variants are not displaced from adhesion sites.