Fig. 4.

Y783A mutations in the β1-integrin tail alter integrin-dependent cell functions. (A,B) Wild-type and Y783A mutant CD cells grown on plastic in the presence of 10% FBS, stained with rhodamine-phalloidin and visualized by immunofluorescent microscopy. Scale bars: 50 μm. (C,D) Wild-type and Y783A mutant CD cells placed in 3D collagen I/Matrigel gels for 7 days in the presence of 5% FBS, stained with rhodamine-phalloidin and visualized by confocal microscopy. Scale bars: 50 μm. (E) CD cell populations were allowed to adhere to collagen I (Col-1) or laminin 511 (Ln-511) for 1 h. Data are mean±s.d. of three experiments in triplicate. *P<0.05 (between wild type and mutant). (F) CD cells were plated on transwells coated with Col-1 or Ln-511, and migration, measured as cells per high powered field (hpf), was evaluated after 4 h. Data are mean±s.d. of three experiments in triplicate. *P<0.05 (between wild type and mutant). (G) CD cell populations were plated on collagen I (Col-1) or laminin 511 (Ln-511). After 24 h, the cells were treated with [3H] thymidine and incubated for a further 24 h. [3H] thymidine incorporation (in cpm) was then determined. Data are mean±s.d. of three experiments in triplicate. *P<0.05 (between wild type and Y785A integrins).