Figure 3

In vivo Chomatin ImmunoPrecipitation (ChIP) analysis for putative TFAP2A binding sites on the core promoters of potential TFAP2A target genes. Pre-cleared chromatin from HeLa or HepG2 cells was immunoprecipitated with either non-specific IgG (IgG, negative control) or anti-RNA polymerase II or anti-acetylhistone H3 (positive controls: Positive Ctrl) or anti-TFAP2A (TFAP2A) antibodies. Immunoprecipitated DNA or non-immunoprecipitated chromatin samples (input) were amplified by PCR using primer pairs designed with the NCBI Primer Designing Tool (see Methods) across the TFAP2A putative binding sites. CD59: CD59 molecule, complement regulatory protein; CXCL1: chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity alpha); DCLBD2/ESDN: endothelial and smooth muscle cell derived neuropilin like molecule; EREG: epiregulin; FASTK: fast kinase; GLO1: glyoxalase I; IFI44: interferon-induced protein 44; PLCXD2: pleckstrin homology-like domain family B member 2; PPARG: peroxisome proliferator activated receptor gamma; SLIT2: slit homolog 2 (Drosophila). C-, PCR negative control; FC, fold changes obtained from the microarray analysis [39]. Number (n°) of TFAP2A binding sites identified computationally. Three independent experiments were performed and a representative one is shown here.