Fig. 9. WAVE2 is recruited to cell-cell contacts through its N-terminus. (A) GFP-tagged WAVE2 constructs were expressed and their localization was examined and quantified in MDCK cells. All data are mean ± s.d. Three independent experiments were performed. WHD, WAVE-homology domain; B, basic domain; PR, proline-rich domain; VCA, verprolin-homology, cofilin-homology and acidic domain. Bar, 15 µm. (B) Pull-down assay. (Left panels) MDCK cell lysates were incubated with GST-tagged truncated WAVE2 proteins and immunoblotted with anti-Abi1 and anti-HSPC300 antibodies. (Right panels) MDCK cells in which GFP-tagged Abi1 or HA-tagged HSPC300 were expressed were lysed and then incubated with GST-tagged full-length WAVE2 proteins and immunoblotted with anti-GFP and anti-HA antibodies, respectively. (C) WHD-GFP was transiently expressed in CTRL, Abi1 or HSPC300 siRNA-treated cells. Expression levels of WHD-GFP were similar in these cells (data not shown). Cells were fixed and the localization of WHD-GFP was examined and quantified. These data indicate that localization of WHD was dependent on Abi1 and HSPC300. Bar, 30 µm. All data are mean ± s.d. of three independent experiments.