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Fig. 6 FIGURE 6. DOC treatment up-regulates the expression of inflammatory genes leading to female-biased population. Zebrafish juveniles were exposed to DOC (200 μm) and analyzed for changes in p65, ptgs2a, and sex ratio. Shown is analysis of p65 levels after 6 days of exposure at 15 dpf (A). 24 h of exposure of 41 dpf juveniles was followed by analysis of p65 (B) and ptgs2a (C) expression in the gonads. Testis explants were exposed to heat-killed bacteria (5 × 107), DOC (200 μm), and NAI (20 nm) for 24 h, and total RNA was extracted followed by qRT-PCR analysis of p65 (D). Zebrafish juveniles at 70 dpf were exposed to heat-killed bacteria (5 × 107) and DOC (300 μm) for 2 days, and gonads were isolated for Western blot analysis (E). Larvae at 15 dpf were exposed to DOC (200 μm) for 20 days, and sex ratio was determined at 70 dpf. The mean and S.D. of two independent experiments are shown. Each group contained at least 30 individuals (F). Statistically significant difference between groups was determined using Student's t test (*, p < 0.05; **, p < 0.01; ***, p < 0.001). Error bars represent the mean ± S.D.