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. 2010 Aug 19;6(8):e1001065. doi: 10.1371/journal.pgen.1001065

Figure 4. Transient transfection analysis of p63 binding sites at SHFM1 locus on chromosome 7.

Figure 4

(A) The p63 binding sites SHFM1-BS1, -BS2 and -BS3 were tested in transient transfection assays in Saos2 cells. Transcription of the luciferase reporter was strongly activated by ΔNp63α through SHFM1-BS1 binding site but only weakly activated through -BS2 and -BS3. (B) Activation was abolished when point mutations were introduced into the p63 binding motif in the SHFM1-BS1 binding site where the essential cytosine and guanine bases were mutated to adenosines. (C) Activation was impaired by p63 EEC mutations R204W, R279H and R304W and slightly reduced by SHFM1 mutation K194E and AEC mutation L517F. (D) No additional activation was observed when SHFM1-BS1, -BS2 and -BS3 were cloned in front of the SV40 promoter driving luciferase. (E) No activation was observed on the mouse Dlx5 promoter (no BS) and when SHFM1-BS1, -BS2 and -BS3 were cloned in front of the Dlx5 promoter.