Figure 2.

LPA activates endogenous RhoA in N1E-115 cells. (A) N1E-115 cells were cultured overnight in serum-free medium and were subsequently stimulated with LPA (1 μM) or bradykinin (1 μM) for the indicated periods. Cell lysates were prepared and incubated with GST-ROK. The activation state of RhoA was then assessed by anti-RhoA Western blotting. (B) RhoA activation depends on tyrosine kinase activity. Cells were pretreated with either genistein (25 μM, 30 min) or tyrphostin 47 (150 μM, 30 min) before stimulation with LPA (3 min). The activation state of RhoA was then assessed as in A.