Fig. 11.

The FLNa-binding domains of SH2B1β, pTyr-PAK1, and FLNa are required for PRL-induced cell migration. A, A7 and M2 cell lines were cotransfected with GFP-PRLR, mRFP-tagged WT SH2B1β or SH2B1β 3Δ mutant, and myc-tagged WT PAK1 or PAK1 Y3F mutant. After deprivation, monolayers of cells were scarified and incubated in deprivation media with or without 400 ng/ml PRL. The percentage wound closure in 8 h was calculated for each condition. Bars represent mean ± se. *, P < 0.05. B, Clones of T47D cells stably expressing GFP, GFP-containing PAK1 WT, or PAK1 Y3F were transiently transfected with vector, WT SH2B1β, or SH2B1β 3Δ mutant. Monolayers of cells were wounded in the presence or absence of 200 ng/ml PRL and assessed after 18 h. Bars represent mean ± se. *, P < 0.05 compared with cells expressing vectors and treated with PRL. Each experiment was repeated three times. n = 30 for each experimental condition. vctr, Vector.