Individual Rac GTPases Mediate Aspects of Prostate Cancer Cell and Bone Marrow Endothelial Cell Interactions
Effect of Rac1 GTPase on β1 integrin activation. (a) Comparison of total, partially and fully activated β1 integrinsin control and Rac1 depleted PC-3 cells as assessed by FACS analysis. Cells were transfected with either siRNA-scrambled control or Rac1 siRNA(2) and left unstimulated or treated with 100 ng/mL CCL2. FACS analysis was performed after incubating fixed cells with the antibodies MAR4 (for total β1 integrin), N29 (for partially active β1 integrin), and HUTS21 (for fully active β1 integrin). (b) is a rescue experiment demonstrating that introduction of either an RNAi-insensitive Rac1 or RhoG GTPase leads to restoration of active β1 integrin levels in CCL2-treated PC-3 cells. Shown are the results of triplicate experiments showing the percentages of 10,000 gated events with significance being *. Capped lines signify a comparison and significance between siRNA-depleted cells and cells rescued with siRNA-insensitive constructs.
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