Fig. 3.

Rac1 ¹⁰⁶PNTP¹⁰⁹ motif is required for its association with PLC-γ1. A, Cos-7 cells were transfected with GFP-tagged wt Rac1 or Rac1PP/AA. The cells were either not treated or treated with EGF (15 min). The transfected Rac1 was immunoprecipitated with GFP antibody, and the resulting immunoprecipitates were immunoblotted with antibodies to GFP and PLC-γ1. B, Quantification of the data from at least three independent experiments as described in panel A. C, The effects of Rac1 ¹⁰⁶PNTP¹⁰⁹ on its association with the PLC-γ1 SH3 domain. COS-7 cells were transfected with GFP-tagged wt Rac1 or Rac1PP/AA. With or without EGF treatment (15 min), the cells were lysed and incubated with GST or GST-fusion PLC-γ1 SH3 domain bound to glutathione-agarose beads. Bound proteins were analyzed by immunoblotting with anti-GFP antibody.