Figure 6.

GPR56-mediated phosphorylation of Src occurs upstream of Fak and is independent of RhoA activation. Western blots showing knockdown of A, G⍺₁₃ and B, G⍺₁₂ or G⍺_q does not significantly affect GPR56-mediated Src/Fak signaling in 293T cells. C and D, western blots showing effect of inhibitors of C, RhoA (Rho Inhibitor I, 2 μg/ml), Src (saracatinib, 10 μM), and D, Fak (defactinib, 10 μM) on constitutive and 10C7-induced phosphorylation in 293T-GPR56 stable cells. Cells were pretreated with inhibitors for 3 h, then treated with 10C7 (3 μg/ml or 20 nM) for 45 min. E, quantification of at least three independent experiments as shown in C and D. Statistical significance for western blots was performed by one-way ANOVA, ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001. Error bars, S.D.