Figure 5
Effect of ERCC1-XPF down-regulation on cisplatin ICL damage and sensitivity. A) MGH-U1 cells were transfected with 10 nM ERCC1 siRNA and analysed 48, 72 and 96 h after transfection for ERCC1 and XPF expression. RPA2 served as loading control. B) MGH-U1 cells were transfected with 10 nM ERCC1 siRNA or control siRNA, cultivated for 96 h, treated with cisplatin (6 μg/ml) for 1 h and post-incubated for 24 or 48 h. γH2AX fluorescence was quantified in MGH-U1 parental cells and cells transfected with either ERCC1 siRNA or control siRNA. For each data point, more than 400 nuclei/experiment were examined, the γH2AX fluorescence was determined and the % cells with a fluorescence intensity more than 500 units (control value) were plotted. Each column represents the mean of 3 independent experiments. C) MGH-U1 cells were transfected with 10 nM ERCC1 siRNA or control siRNA, cultivated for 96 h, treated with cisplatin (9 μg/ml) for 1 h and post-incubated for 48 or 72 h. Apoptosis was measured by flow cytometry (sub-G1 content). * represents statistical significance.