Figure 5.

p190-induced phenotypes are Rho dependent. (A) Diagram of the p190RhoGAP mutants: Full-length (FL), ΔGAP, ΔGBD, Middle Domain (MD), dominant negative GAP inactive R1283A, and GAP-only domain (GAP). (B) Western blot analysis of HA-p190RhoGAP mutants: MDA-MB-468 cells were transfected with plasmids encoding the indicated mutants of p190, and 48 hours later, cell lysates were immunoblotted with anti-HA antibody. (C) Effect of p190 mutations on induction of apoptosis. Indicated cells were transfected and analyzed 48 hours later as in Figure 1. Results are expressed as the mean percentage of p190-overexpressing cells that were TUNEL positive ± SEM (n > 3). (D) Rescue of p190-induced phenotypes by CARho. Indicated cells were cotransfected with equivalent molar amounts of p190 and CARho (Q63L) or with p190RhoGAP alone and cultured for 48 hours. Where indicated, only cells expressing FLp190 alone or both FLp190 and CARho were assessed for apoptosis, multinucleation, or dendrite-like formation. Results are expressed as the mean percentage of HA-p190– or HA-p190/CARho–expressing cells that exhibited the indicated phenotype ± SEM (n > 3). Statistical significance is determined within individual cell lines. *P < 0.005 or **P < 0.05, comparing presence of CARho to its absence in p190-overexpressing cells.