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Fig. 6. Roscovitine inhibits Trio-GEF activity. (A) hEK293 cells stably expressing PAM-1 were transiently transfected with vector, myc-tagged Trio (Trio) or Myc-tagged Trio-GEF1 (T-GEF1). Control cells (-) and cells pre-treated with roscovitine (+) were extracted and aliquots were probed for Myc-tagged products (top) and total Rac (center); activated Rac was assessed following adsorption to GST-Pak-CRIB bound to glutathione agarose (bottom). A representative experiment is shown. (B) Rac activation was quantified from three independent assays; error bars show standard deviation.