ILK-1–dependent recruitment of Dvl is necessary for BMP-2–mediated hPASMC migration. (A) Confocal microscopy was used to localize ILK-1 (red) and Dvl (green) in hPASMCs stimulated with 10 ng/ml BMP-2. DAPI (blue) was used to label the nuclei. Bars, 10 µm. (B) Co-IP of V5-tagged ILK-1 and WT, ΔDEP, and ΔPRS Dvl-GFP constructs under 10 ng/ml BMP-2 stimulation. Levels of V5-tagged ILK-1 were measured against levels of α-tubulin in whole cell lysates analyzed in a separate gel. (C) RhoA (left) and Rac1 (right) activation in BMP-2–stimulated hPASMCs transfected with either WT, ΔDEP, or ΔPRS Dvl was performed as described in Materials and methods. Levels of active RhoA and Rac1 were measured against levels of total RhoA and Rac1 in whole cell lysates analyzed in a separate gel. (D) Boyden chamber assays were performed as previously described to measure the impact of WT, ΔDEP, and ΔPRS Dvl on BMP-2–induced hPASMC motility. Bars represent means ± SEM from n = 3. **, P < 0.001; and ***, P < 0.0001 versus time 0 in B and C using one-way ANOVA with Dunnett’s. ***, P < 0.0001 versus control (CON) unstimulated; and ^###, P < 0.0001 versus WT stimulation in D was determined using one-way ANOVA with Bonferroni’s. IP, immunoprecipitation. IB, immunoblot.