Fig. 3. Polarization of B16-F1 cells expressing vinculin variants. Populations expressing vinculin constructs were fixed 6 hours after seeding onto laminin and actin filaments stained using phalloidin. (A,B) Representative cells of populations expressing vinculin-wt (A) or vinculin-LD (B) are shown in fluorescence images (left) and as outlines of cells (right). Classification of cells into the categories polarized (P), unpolarized (N) and ambiguous (A) was based on cell shape and cytoskeletal staining. Scale bar, 10 µm. (C) Proportions of cells in each category taken from >600 cells (n=3 independent experiments) for each vinculin construct. Notice the significant difference between the numbers of polarized motile cells in the vinculin-wt-expressing B16-F1 and vinculin-LD-expressing cells.