Hck interacts with YAP and requirement of YAP for transactivation of Ipaf by p73. (A) Cos1 cells transiently expressing GFP-YAP with Hck were immunoprecipitated with control rabbit IgG (C) or Hck polyclonal antibody. The immunoprecipitates were resolved by SDS-PAGE and subjected to western blotting to detect YAP and Hck. (B) Extract of Cos1 cells transfected with GFP-YAP was incubated with GST, GST-SH3Hck, GST-mSH3-Hck recombinant proteins bound to Glutathione Sepharose beads. Proteins bound to these beads were immunoblotted with GFP and GST antibodies. (C) YAP-shRNA downregulates GFP-YAP expression. Hela cells transfected with GFP-YAP (50 ng) with either control shRNA I and II (200 ng) or YAPshRNA I, II and III (200 ng) subjected to immunoblotting with anti-GFP antibody. GFP (50 ng) was used as transfection efficiency control. (D). Effect of YAP shRNA on p73α mediated Ipaf promoter transactivation. HeLa cells transiently transfected with YAPshRNA I, II (200 ng) or control shRNA I (200 ng) in the presence or absence of p73α (2 ng) along with Ipaf-CAT promoter construct (150 ng) and pCMV-β Gal (50 ng) were subjected to CAT activity measurements.