Figure 3

Myc and Max are associated with the Aurora-A promoter. A. Schematic representation of the potential Myc binding sites of the Aurora-A promoter. B. LS174T cells were treated or not with doxycyclin, soluble chromatin was immunoprecipitated with anti-Myc or anti-acetylated-H3 polyclonal antibodies and DNA samples were then amplified using primers that cover the -668/-400 region of the Aurora-A promoter. IgG immunoprecipitations were used as controls (n = 3 +/- sd). C. HCT116 were synchronized in G1/S with hydroxyurea and released for the indicated times in growth medium complemented with 3% serum. Soluble chromatin was immunoprecipitated with anti-Myc or anti-Max antibodies and DNA samples were then amplified using primers that cover the -668/-400 region of the Aurora-A promoter and quantified as compared to IgG immunoprecipitations (n = 3 +/- sd). D, E. The association of Myc and Max on the Aurora-A promoter was analyzed by a serial ChIP experiment. HCT116 were synchronized as described above, the soluble chromatin was immunoprecipitated with Myc antibodies, immune complexes were released and reimmunoprecipitated with IgG or Max antibodies. DNA samples were then amplified using primers that cover the -668/-400 region of the Aurora-A promoter and analyzed by semi-quantitative PCR (D) or quantitative PCR (E, n = 3 +/- sd).