Skip to main content
. Author manuscript; available in PMC: 2012 Mar 25.
Published in final edited form as: Biochem Biophys Res Commun. 2011 Feb 23;406(4):574–579. doi: 10.1016/j.bbrc.2011.02.093

Figure 1. GEFH1 binds to ASAP1 in vivo.

Figure 1

FLAG-tagged BAR-PH was expressed in HEK293 cells and immunoprecipitated with FLAG antibody conjugated beads. GEFH1 was detected in the precipitate by immunoblotting with an anti-GEFH1 antibody. (B) NIH 3T3 cells were transfected with expression vectors for active Src or GFP. Proteins from the cell lysates were precipitated with anti-GEFH1 antibody and ASAP1 was detected in the precipitate by immunoblotting. (C) EE-tagged ASAP1 was expressed in NIH 3T3 fibroblasts and was immunoprecipitated with anti-EE antibody. GEFH1 was detected in the precipitate by immunoblotting. (D) The recombinant GEFH1 proteins used in this paper. The two-hybrid clones obtained were indicated at the top as prey. CC; coiled-coil, Zn; Zinc finger, DH; Dbl-homology, PH; Plecstrin homology domain. (E) Flag-tagged [1-438]ASAP1 (BAR-PH) was expressed in NIH 3T3 fibroblasts with full length GFP-tagged GEFH1 or [1-572]GEFH1 (ΔCC GEFH1) and proteins were immunoprecipitated with either nonspecific IgG or an anti-Flag antibody. GEFH1 was detected by immunoblotting. (F) EE-tagged full length ASAP1 was coexpressed with either full length GEFH1 or ΔCC GEFH1. Proteins were immunoprecipitated with a nonspecific IgG or with an antibody to the EE epitope tag. GEFH1 was detected by anti-GFP antibody.