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Figure 2


Fig. 2. p63 and Dlx3 expression in primary mouse keratinocytes cultured in vitro. Dlx3 (A) and TAp63 (B) mRNAs were induced, and {Delta}Np63 (C) mRNA was downregulated after 12- and 24-hours of high-[Ca+2] treatment. (D-F) p63 binds to the Dlx3 promoter region in vitro and in vivo. (D) EMSA assay performed with a DNA fragment that included the Dlx3 p63-binding site 1 and 2 using nuclear extract from primary keratinocytes (NE), and in the presence of 100 M excess of specific (SC) and nonspecific (NC) competitors. (E-F) ChIP analysis on mouse keratinocytes with either control IgG or p63 antibody (p63 Ab) on the region of the Dlx3 promoter containing the p63-binding sites by regular (E) and real-time (F) PCR. (G) ChIP analysis on TAp63{alpha}-transfected H1299 cells with no antibody (no Ab) or p63 antibody on DLX3 and JAG2 promoters. ACHR was used as a control.