Figure 3.

Rho GTPase activity assays. Lysates from NMuMG-NYPD/vector or NMuMG-NYPD/p110 cells were incubated with GST fusion protein containing the binding domain of Rac1 or Cdc42 and bound to Glutathione Sepharose beads. Active Rac1 and Cdc42 specifically bound to the beads were eluted in Laemmli sample buffer and analyzed by western blotting using anti-Rac1 or anti-Cdc42 antibodies. The amount of total protein was determined by analysis of an aliquot of total lysate, and equal loading was verified with actin.