Constitutively active βC induces FN production, RhoA-Rac1, motility, and proliferation in hPASMCs. (A) Levels of secreted FN were measured in hPASMCs transfected with either vector or ΔβC. Densitometry values were expressed as the ratio of OD of FN relative to the Ponceau stain of an unrelated band in media samples concentrated by ultrafiltration. (B) RhoA (left) and Rac1 (right) activation in BMP-2–stimulated hPASMCs transfected with either vector or ΔβC 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. (C) Boyden chamber assays were performed as previously described in this paper to measure the impact of vector or ΔβC on hPASMC motility. (D) Cell count studies in hPASMCs transfected with either vector or ΔβC were performed for 72 h. Bars represent means ± SEM from n = 3. ***, P < 0.0001 versus vector using an unpaired t test.