Fig. 3.

ATM activation and an E2F1 binding sequence were required for activation of BMCC1 promoter in NB cells with DNA damage. a Schematic representation of BMCC1 promoter constructs [6]. Vertical line indicated the position of E2F-binding sites. The E2F-binding site with X-mark harbors one base mutation to diminish the consensus sequence. b CDDP-mediated activation of BMCC1 promoter is suppressed by ATM inhibitor treatment. pGL3-BMCC1-luc and pRL-TK (Renilla luciferase reporter plasmid) transfected SK-N-AS cells were maintained in normal medium for 46 h and then transferred in a fresh medium containing the appropriate ratios of CDDP and ATM inhibitor. Two hours after treatment, the luciferase activity was measured for all samples. Firefly luciferase activity was normalized to that of Renilla luciferase. Mean values were calculated from triplicate experiments. Error bars indicate standard deviation (*P < 0.01, n = 3). c The putative E2F-binding site S9 is responsible for the CDDP-dependent activation of BMCC1 promoter. SK-N-AS cells were transfected with the luciferase reporter plasmids. Forty-six hours after transfection, the cells were cultured with or without CDDP. Two hours after treatment, luciferase activity was measured for all samples (*P < 0.001, n = 3). n.s. stands for not significant