Figure 4
From: Rac inhibits thrombin-induced Rho activation: evidence of a Pak-dependent GTPase crosstalk
Pak1 binds the DH-PH domain of P115RhoGEF. (A) Constructs used in these experiments. (B) HEK293T cells were transfected with expression plasmids encoding the AU1-tagged constructs shown in (A) in the presence or absence of wild-type myc-tagged Pak1 and/or the activated Rac mutant RacQL (Rac). Lysates were immunoprecipitated with anti-myc antibody, and immunoprecipitates tested for GEF content by western blotting with anti-AU1 antibody. (C) HEK293T cells were transfected with expression plasmids encoding AU1-tagged DH-PH domains of P115RhoGEF (P115), PDZ-RhoGEF (PRG), and LARG (LARG) in the presence or absence of wild-type myc tagged Pak1 and/or the activated Rac mutant RacQL (Rac). The P115RhoGEF ΔN construct is included as a control. Lysates were immunoprecipitated with anti-myc antibody, and immunoprecipitates tested for GEF content by western blotting with anti-AU1 antibody.