Figure 4.

EE₂ modulates the expression of genes coding for immune-relevant molecules in the gonad. Specimens at both pre-SG and SG stage were treated with 0 and 5 µg EE₂/g food during 7 and 28 days. Afterwards, the mRNA levels of il1b (a); tnfa (b); tgfb1 (c); mmp9 (d); mmp13 (e); tlr9 (f); and mhc1a (g) were determined in the gonad by real-time RT-PCR. Total RNA was obtained after pooling the same amount of mRNA from five to six fish/group. Data represent means ± SEM of triplicates of the same pooled sample. The asterisks denote statistically significant differences after Student t test between: (i) the untreated control group and the EE₂ treated group at each time point and spermatogenic condition and (ii) the untreated control groups of the two spermatogenic conditions within the same sampling date. * p < 0.05; ** p < 0.01 and *** p < 0.001. ns, not significant.