Figure 4. The effects of genistein on mitochondrial membrane potential, ROS levels and the activation of DNA damage response kinases in Eca-109 cells. (A, B) Mitochondrial membrane potential in Eca-109 cells treated with genistein for 24 h was detected through flow cytometry. ROS levels in Eca-109 cells treated with genistein for (C, D) 24 h or (E, F) 48 h were detected through flow cytometry. (G, H) The ROS levels in Eca-109 cells treated with 5 mM NAC for 6 h, followed by genistein treatment for 48 h. (I) The mRNA levels of γH2AX, ATM, ATR and CHK2 in xenograft tumor tissues were measured through qPCR (n = 6 per group). (J) The mRNA levels of γH2AX, ATM, ATR and CHK2 were analyzed through qPCR. (K, L) The protein expression of γH2AX, ATM, ATR, CHK2, p-ATM, p-ATR and p-CHK2 in Eca-109 cells treated with genistein for 72 h was measured through western blotting. All experiments were independently repeated three times. Data are analyzed using one-way ANOVA with Dunnett’s test and are presented as the mean ± SD. *P<0.05; **P<0.01; ***P<0.001; NAC, N-acetylcysteine; Gen, genistein.