Fig. 6.

PRL-1 activity is interrupted by Y-27632. A, A549 cells stably expressing PRL-1 (WT) were cultured in BME with 10% FBS with either 0.2% DMSO alone or 5 or 15 μM Y-27632 for 12 h. The cellular extracts were analyzed by Western blotting using Myc-Tag PRL-1, RhoA, Cdc42, Rac, and GAPDH as indicated. The expression levels of GTP-RhoA were assessed using a Rho activation assay kit. GAPDH was used as a loading control. B, PRL-1 with Y-27632 inhibits cell migration in the scratch wound-healing assay and cell invasion using Matrigel invasion chambers. ∗∗, P < 0.01 compared with WT. C–E, PRL-1 effects on cell shape and actin stress fiber after treatment with Y-27632. A549 cells stably expressing WT PRL-1 were cultured in BME with 10% FBS for 24 h. The cells were incubated in a 5% CO₂ incubator with either vehicle alone (C), 5 μM Y-27632 for 30 min (D), or 5 μM Y-27632 for 3 h at 37°C (E). The cells were then stained for actin stress fibers.