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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2009 Apr 2;29(6):921–928. doi: 10.1161/ATVBAHA.109.187229

Figure 5. Myocardin, SRF and Brg1 form a complex in vivo and Brg1 binds directly to myocardin in vitro.

Figure 5

A. COS cells were transduced with HA-tagged myocardin and HA-tagged SRF adenovirus. After 24 hours, nuclear protein was harvested and proteins were immunoprecipitated using Brg1, SRF or control IgG antibodies, as indicated. Immunoprecipitated proteins were identified by Western blotting using antibodies against Brg1 or the HA-epitope tags on myocardin and SRF as indicated at the right of the blot. B. A10 SMCs were transduced with omni-tagged myocardin for 24 hours. Nuclear protein was harvested and subsequently immunoprecipitated using myocardin or control IgG antibody. C. SRF, myocardin, and Brg1 or Brm (D) were transcribed and translated in vitro, the expressed proteins were then incubated together (SRF+Brg1/Brm or myocardin +Brg1/Brm) as indicated at the top of the blots. Protein mixtures were immunoprecipitated with myocardin, SRF or IgG control antibodies, as indicated below the blots. The precipitated proteins were analyzed by Western blotting, using antibodies indicated at the right of the blots. On all blots ‘input’ lanes represent 10% of the inputs that were mixed together and used for immunoprecipitates.