Figure 6

Overexpression of HMGA1 correlates with the activation of DNA-damage response. (A) Cells were treated with 50 μM of 5-fluorouracil (left panel) or with 500 nM of doxorubicin (right panel) and collected at the indicated time points after drug treatment. Protein extracts were analyzed western blot to evaluate ATM and H2AX phosphorylation. Total ATM levels are also reported. For H2AX loading control see the β-actin panels in Figure 4. (B) Cell viability curve of GEO CR and FRO cells pretreated or not with 10 μM of KU-55933 for 1 hour, and then with increasing doses of 5FU or doxorubicin. Values are the mean ± S.D. of three experiments performed in triplicate. (C) GEO CR pLKO.1 and GEO CR shHMGA1 cells were treated with 1 μM of CTX, collected after 0, 18 and 24 hours of treatment and processed for comet assay. Comets were stained with SYBR Green, visualized by fluorescence microscopy and analyzed by COMET Score software. A representative comet for each experimental point is shown in the left panel. Right panel shows the quantitative analysis using the COMET Score software. *p < 0.05 significance level compared with GEO CR pLKO.1.