Research Paper Volume 2, Issue 12 pp 959—968

Figure 2. Binding of the p-ΔNp63α protein to the ATM promoter in vivo. Wild type ΔNp63α cells (left panels) and ΔNp63α-S385G cells (right panels) were exposed to a control medium and 10μg/ml cisplatin for 24h. (A) ChIP assay of a specific region of the ATM promoter with anti-p-ΔNp63a antibody and anti-DNp63 antibody. As negative controls, we used ChIP of the ATM promoter specific region with rabbit immunoglobulins (IgG) and ChIP of the ATM promoter non-specific region with anti-p-ΔNp63α antibody as indicated. (B) Luciferase reporter assay. Both types of cells were transfected with 100 ng of the promoter-less pGL3 plasmid or pGL3-ATM (1259bp) promoter plasmid along with 1 ng of the pRL-SV40 plasmid for 24h. Cells were also transfected with 100 ng of the ΔNp63α-FL (Flag) or ΔNp63α-S385G-FL expression cassettes, as indicated. Cells were exposed to control medium (Con) and 10 μg/ml cisplatin (CIS) for 24h. Luciferase reporter assays were conducted in triplicate (± SD are indicated, p<0.05) as described in the Materials and methods. Firefly luciferase activity values were normalized by Renilla luciferase values.