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5 ml phenol red-free DMEM for 30 min at 37��C in the dark. The cells were washed, suspended in 0.5 ml ice cold PBS and immediately analyzed for DCF fluorescence using the BD Accuri C6 flow cytometer. In each sample, 10 000 events were counted and the results were presented as fold increase of the fluorescence intensity compared with the control sample. Statistics In all figures the data are expressed as mean �� standard error of mean (SEM). Statistical analyses were performed by using STATISTICA 7.0 (StatSoft Inc.). The effect of different doses of MEL and DMEL either in the presence or absence of diquat on cell viability was analyzed by two-way analysis of variance (ANOVA). The effect of different doses of diquat in the presence or absence of melatonin on ROS production of HeLa cells was analyzed with two-way repeated measures ANOVA. The effect of different doses of melatonin in the presence of diquat on ROS production was evaluated with one-way repeated measures selleck inhibitor ANOVA. Differences between individual groups were compared by Fisher's least-squares difference post hoc tests. Results Diquat negatively influenced viability of Vero cells in a dose-dependent manner (F (3,21)=6.16, pTalazoparib ic50 by an interaction between diquat and MEL (F (3,41)=7.94, pbinedaline capacity of these compounds against negative effects of diquat. On the contrary, the viability of cells treated with the highest concentration of MEL was significantly lower than that of the control. Such an inhibitory effect was seen after administration of DMEL in an even 10-times lower concentration compared with MEL. Figure 1 Viability of Vero cells treated with different doses of (A) melatonin (MEL) and (B) dihydromelatonin (DMEL) in the presence or absence of 10 ��M diquat (DIQ) for 24 h. Cell viability was evaluated by MTT test. Data are presented as mean �� ... Table 1 Effect of different doses of diquat on viability of Vero and HeLa cells after incubation for 24 h and assessed by the MTT test.