Fig. 8. Focal adhesion formation and cell adhesion are downregulated in 3T3-L1 cells overexpressing ADAM12 at the cell surface. Cells (suspended in A,B and adherent in D-J) with constitutive expression of ADAM12-cyt at the cell surface (A,D,G) and control cells (B,E,H) were examined. (A-C) ADAM12 immunostaining and western blot analysis. (D-F) Staining with rhodamine-conjugated phalloidin to monitor F-actin organization (D,E) showed that ADAM12-cyt-expressing cells were smaller than control cells, and that, compared with the control cells, a significantly higher percentage of cells had fewer than five stress fibers (P<0.02) (F). (G-I) Vinculin immunostaining (G,H) showed that a higher percentage of ADAM12-cyt expressing cells than control cells had fewer than five focal adhesions (P<0.02) (I). (J) Compared with the control cells, cell attachment on fibronectin was reduced (P<0.01) in ADAM12-cyt-expressing cells. Bars, 10 µm (A,B) and 15 µm (D,E,G,H).