Figure 6

In vivo Chomatin ImmunoPrecipitation (ChIP) analysis for putative SP1 binding sites on the core promoters of TFAP2A target genes. Pre-cleared chromatin from HeLa cells was immunoprecipitated with either non-specific IgGs (IgG, negative control) or anti-RNA polymerase II or anti-acetylhistone H3 (positive controls: Positive Ctrl) or anti-SP1 (SP1) or anti-TFAP2A (TFAP2A) antibodies (Ab). 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 or the SP1 putative binding sites. ADAMTS1: ADAM metallopeptidase with thrombospondin type 1 motif, 1; CASP9: caspase 9; KRT16: keratin 16; KRT17: keratin 17; PLCXD2: pleckstrin homology-like domain family B member 2; TGFBI: Transforming Growth Factor Beta-Induced. The number (n°) of computationally predicted high score TFAP2A or SP1 binding sites are indicated. C-, PCR negative control; FC, fold changes obtained from the microarray analysis [39]. Three independent experiments were performed and a representative one is shown here.