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. Author manuscript; available in PMC: 2007 Mar 2.
Published in final edited form as: Biochemistry. 2006 Oct 3;45(39):11727–11736. doi: 10.1021/bi061180d

Figure 5.

Figure 5

Cl-amidine inhibits the PAD4-mediated enhancement of the p300GBD–GRIP1 interaction in CV-1 cells. (A) Schematic depiction of the mammalian two-hybrid assay used to evaluate the efficiency of the p300GBD–GRIP1 interaction. (B) CV-1 cells were transfected with a luciferase reporter construct as well as plasmids encoding the proteins depicted in this panel. The indicated concentrations of Cl-amidine were added to the cell culture medium and the mixtures incubated for 40 h. Cell extracts were then prepared, and the luciferase activity present in these extracts was quantified. The catalytically defective PAD4 C645S mutant was also transfected into this system to demonstrate that the decrease in the strength of the p300GBD–GRIP1 interaction is not a nonspecific effect.