AFAP-110 is overexpressed in prostate cancer and contributes to tumorigenic growth by regulating focal contacts
J. Clin. Invest. Jing Zhang, et al. 117:2962 doi:10.1172/JCI30710 [Go to this article.]

Figure 6
Restoration of integrin β1 expression and focal adhesions by ectopic expression of wild-type AFAP-110 and functional mutant variants. (A) Schematic representation of wild-type AFAP-110 and functional mutant variants. Deletion of the PH1 domain (AFAP-110 Δ180-226) abolishes the association of AFAP-110 with PKC. A single amino acid mutation that changed a proline residue to an alanine (AFAP71A) at the SH3-binding motif abrogates the ability of AFAP-110 to interact with Src. (B) Immunoblotting with antibodies against AFAP-110 and integrin β1. Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants was visualized as bands that localized slightly higher than endogenous AFAP-110 on the membrane. Vinculin expression was used as a loading control. (C) Immunofluorescence staining of vinculin was performed using a monoclonal anti-vinculin primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Images were converted to grayscale for best visualization of focal adhesion structures (yellow arrows). Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants in AFAP-110–downregulated clone 309 were visualized as proteins emitting green fluorescence under microscope (green). Representative images are shown; scale bars: 20 μm.