FIGURE 2.

Contractile stimulation promotes Abi1/N‐WASP coupling and translocation. A, Input or Abi1 precipitates of untreated (UT) or ACh‐treated HASM cells were immunoblotted with antibodies against Abi1 or N‐WASP. ACh treatment increases the ratios of N‐WASP/Abi1 immunoprecipitates in cells. Data are mean values of experiments from four batches of cell culture. Error bars indicate SE. B and C, Representative micrographs illustrating the effects of ACh (10⁻⁴ M, 5 minutes) on the spatial localization of Abi1 and pN‐WASP or N‐WASP in HASM cells. The arrows indicate a single line scan to quantify the fluorescent signals in cells. The line scan graphs show relative fluorescent intensity. Black/white images show the shape and edge of these cells. Image analysis for protein localization was described in the section of Materials and Methods. Scale bar, 10 µm. Protein distribution in cells is expressed a ratio of the intensity at the cell periphery to cell interior. Data are mean values of experiments from three batches of cell culture. Error bars indicate SE. **P < .01