FIGURE 6.

Identification of AP-1 components binding to the new distal AP-1 element within the TF promoter. Chromatin immunoprecipitation assay and electrophoretic mobility shift assay demonstrate specific binding of c-Jun, Fra-1, and ATF-2 to the new AP-1 site in response to P2Y2R activation in HCAEC. A–C, ChIP assay of the new AP-1 site in the TF promoter with anti-c-Jun, Fra-1, and ATF-2 antibodies using HCAEC chromatin. Histograms were acquired from four independent real time PCR experiments followed by electrophoresis gel imaging of amplified PCR products. D–F, confirmation of the roles of c-Jun, Fra-1, and ATF-2 binding to the new AP-1 site with EMSA experiments. 1 and 2 μg of antibody were used in binding reactions or 2 μg of IgG was used as negative controls. Location of arrows indicate band of interest. Data were representative of three independent experiments. IP, immunoprecipitation.