BMP-2 recruits the PDZ and DEP domains of Dvl to activate RhoA and Rac1 and induce hPAEC motility. (A) Diagram illustrating the structure of the four Dvl-GFP constructs used in the experiments described in this study. (B and C) Dvl constructs illustrated in A were individually nucleofected in hPAECs to assess impact on RhoA (B) and Rac1 (C) activation in the presence of 10 ng/ml BMP-2. Pull-down experiments were performed and analyzed as in Fig. 5. Error bars denote mean ± SEM for three different experiments with triplicate assessments. *, P < 0.01 and **, P < 0.001 (vs. control [CON]). (D) Microcarrier bead migration assay of hPAECs nucleofected with Dvl constructs and exposed to BMP-2 as in A. ***, P < 0.0001 (vs. WT and ΔDIX unstimulated). (E) Representative confocal images of hPAECs nucleofected with GFP-tagged Dvl constructs. Cells were starved for 24 h and incubated with BMP-2 as in B. Actin was labeled with Alexa Fluor 555–phalloidin (red), and nuclei were stained with DAPI (blue). Quantification of Dvl distribution in the cytoplasm and the periphery was performed as described in Materials and methods. ***, P < 0.0001 (vs. baseline). (D and E) Error bars denote mean ± SEM for three different experiments with quadruplicate assessments. Bars, 10 μm.