Male zebrafish were treated for different periods as indicated. After treatment, male zebrafish were paired with females with induced ovulation. (A) The behavior of only chasing was scored as 0.5, and chasing and inducing spawning was scored as 1.0. (B) The total egg number obtained by mating was compared among treatments. (C) Survival rate of fertilized eggs obtained by mating. Each value represents the mean of data from three or more trials with three different males (n≧9). Vertical lines indicate standard deviation. One-way ANOVA was used to assess significant differences among the treated groups. Different letters represent significant differences among the data. b, P value < 0.05, c, P value < 0.01, d, P value < 0.001, e, P value < 0.0001. The mark ‘a’, which indicates that the group did not show significant differences from the control (ethanol, EtOH), is omitted. All the low data represented in the figure is available in S1 Table, also data of statistical analysis is available in S2 Table.