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A) EMSA reveals binding of ΔNp63α in nuclear extracts of HaCaT cells using a radiolabeled probe corresponding to Hs II of the human K5 gene. Competition assays show that the KSC oligonucleotide, which contains a previously identified ΔNp63α binding site within the K14 gene, competes with binding of Hs II probe (lanes 2–3). A p53 consensus (lanes 4–5) and a WT oligonucleotide (lane 6–7) also compete for binding while a MT oligonucleotide did not affect Hs II binding (lanes 8–9). The highest complex (asterix) is supershifted (arrow) with the addition of antibodies raised against various domains of ΔNp63 whereas anti-TA antibodies have no effect (lanes 10–13). The lower complexes are likely to be non-specific. B) In vivo occupancy of ΔNp63α to the mouse and human K5 gene. ChIP was performed on HaCaT or primary mouse keratinocytes using two separate antibodies recognizing ΔNp63α as well as a nonspecific IgG as indicated (left panel). Input represents PCR amplification of 1% of the genomic DNA. Primers corresponding to a region of the GAPDH gene serves as a negative control. Right panel shows results obtained from real-time PCR experiments.