FIGURE 4.

Daam1 binds to Fascin via FH domains. A, The lysates of MCF‐7 cells transiently expressing 3FLAG‐Daam1‐full, 3FLAG‐GBD‐FH1, 3FLAG‐FH2 and 3FLAG‐FH2‐DAD were mixed with bacterially expressed His‐Fascin and examined by immunoblotting with anti‐FLAG antibody. B‐D, The morphology of actin filaments in MCF‐7 cells expressed with the indicated constructs was visualized by immunofluorescence microscopy. The number of actin filaments (length > 10 μm) per cell and the length of the longest actin filaments were counted. Bar, 5 μm. E and F, MDA‐MD‐231 cells transfected with the indicated constructs migrated for 6 h in Boyden chambers. The number of migratory cells per field was counted. Bar, 10 μm. Objective lens, magnification, ×20. G and H, MDA‐MD‐231 cells were transiently transfected with the indicated constructs. Pseudopodia (invadopodia) were allowed to extend in 3.0 μm porous Boyden chambers. Invadopodia on the lower side of the membrane were incubated with cortactin antibody and counted per field of microscope. Bar, 10 μm. Objective lens, magnification, ×40; numerical aperture, 0.95. *P < .05. ***P < .001. ****P < .0001