Figure 3.

Dynamin mutants influence cell spreading but not cell adhesion. Rat1 fibroblasts overexpressing dynamin-2 constructs were serum starved for 16 h, replated on FN as indicated and stained for overexpressed dynamin and F-actin. (A) Cell spreading time course on FN-coated coverslips (15 μg/ml coating solution). Noninfected Rat1 cells were used as control (n >100 cells for each data point). (B) Cells spreading for 20 min as in A, stained with rhodamine-phallodin and anti-HA for overexpressed dynamin proteins. Bars, 20 μm. (C) FN dose-response in cell spreading. (D) Effect of dynamin mutants on cell adhesion. Adherent cells were counted (n > 100 for each data point) and the fraction of cells overexpressing dynamin constructs was determined. Results are representative of three experiments (A-D). (E) Comparison of dynamin constructs in cell spreading on FN (15 μg/ml coating solution) for 30 min. Noninfected Rat1 cells served as control. Results are means ± SEM from three experiments.