FIG. 8.
Mapping of the regions of HDAC5 that interact with myocardin. (A) Schematic diagram of HDAC5 and the mutant forms used to map the myocardin-binding domain. (B) Coimmunoprecipitation assays. COS cells were transiently transfected with expression vectors encoding Myc-tagged HDAC5 deletion mutant proteins and FLAG-tagged myocardin. Myc-tagged HDAC5 deletion mutant proteins were immunoprecipitated (IP) from cell lysates with a polyclonal anti-Myc antibody, and coimmunoprecipitating myocardin was detected by immunoblotting (IB) with a monoclonal anti-FLAG antibody (top parts). The membrane was reprobed with anti-Myc antibody to reveal the total amount of Myc-tagged HDAC5 proteins (bottom parts). (C) COS cells were transiently transfected with expression vectors encoding myocardin and HDAC5 deletion mutant proteins and the SM22 luciferase reporter.