Individual Rac GTPases Mediate Aspects of Prostate Cancer Cell and Bone Marrow Endothelial Cell Interactions
Figure 2
Effect of Rac depletion on tumor cell diapedesis across a BMEC monolayer. (a) PC-3 cells were treated with 100 ng/mL CCL2 in a diapedesis assay. Control untransfected (UT) and siRNA control (siScr) cells demonstrated increased diapedesis compared with untreated/untransfected (UN/UT) PC-3 cells. The ability of cells to undergo CCL2-stimulated diapedesis after depletion of Rac1, RhoG, or treatment with iRac was compared to UT and siScr. Rescue experiments of RhoG-depleted cells were performed by the introduction of a siRNA-insensitive RhoG. Rac1-depleted cells were rescued with the introduction of fast cycling RhoG (RhoGQ63L). (b) Depletion of RhoG led to a decrease of total Rac activation in PC-3 cells treated with 100 ng/mL CCL2. Rescue experiments were performed by introducing a siRNA-insensitive RhoG GTPase. Shown are means ± S.D. of at least triplicate analysis representing individual transfections, with significance being ; (*) signifies a significant difference between siRNA-transfected cells and stimulated controls, while (∧) signifies a significant difference between siRNA-transfected and -rescued cells.
