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. 2004 Feb 12;23(4):790–799. doi: 10.1038/sj.emboj.7600073

Figure 5.

Figure 5

Activated Src family kinases contribute to the YAP–Runx2 interaction. (A) ROS 17/2.8 cells, transiently transfected with XPR-YAP and antibodies, were used to immunoprecipitate endogenous Yes or Src tyrosine kinases. Association of YAP with these kinases was assessed by western blotting using HRP-conjugated anti-XPR antibody. (B) ROS 17/2.8 cells, expressing indicated constructs, were treated with 5 μM of PP2 for 1 h and processed for immunoprecipitation. The Runx2–YAP interaction is completely abrogated in the presence of PP2 (panel 1). Panel 2 shows the efficiency of Runx2 immunoprecipitation, while panel 3 shows comparable levels of YAP overexpression in all lanes. (C) As in (B), but antibodies were used to immunoprecipitate endogenous Runx2. The effect of Src DN on the YAP–Runx2 interaction was assessed by western blotting using the HRP-conjugated anti-XPR antibody (panel 1). A mouse monoclonal antibody against Runx2 was used to assess the efficiency of immunoprecipitation of the endogenous Runx2 protein (panel 2). Expression of Src DN and XPR-YAP was confirmed with Src and HRP-Xpress antibodies, respectively (panels 3 and 4). In each immunoprecipitation, appropriate normal IgG was used as a control.