Expression of PARP-1 and γH2AX in hydroxytyrosol treated melanoma cells. A375 cells (A–D) were treated with 250 μM, 375 μM, and 500 μM of hydroxytyrosol and HT-144 cells (E–H) were treated with 250 μM, 350 μM, and 450 μM of hydroxytyrosol for 24 h and 48 h. The expression of PARP-1 (A,E, upper panels, B,F), of the cleaved form of PARP-1 (A,E, upper panels, C,G), and of γH2AX (A,E, middle panels, D,H), was analysed in A375 and in HT-144 cell lines in response to the treatment with hydroxytyrosol through western blot experiments. The equal protein loading was confirmed by the analysis of β-actin expression (A,E, lower panels). The data are representative of several western blot experiments. The error bars indicate the standard deviation. Statistical significance was analysed by Student’s t-test: * p < 0.05 was considered significant; ** p < 0.01 highly significant; *** p < 0.001 very highly significant.