Fig. 4.

Relative aromatase (cyp19a1a) gene expression in ovaries (primary oocyte stage). Relative gene expression for each treatment combination (A) and log₁₀ relative gene expression by individual treatment (B) for n = 6 females per treatment. Relative gene expression = (E ref)^{Ct ref}/(E target)^{Ct target}, where ref is the housekeeping gene (rpl8), target is the gene of interest (cyp19a1a), E is PCR amplification efficiency, and Ct is cycle threshold (number of temperature cycles yielding above-background expression) for that particular gene. High-level clotrimazole exposure (8 μg/L) was shown to be a reliable predictor (P = 0.0003) of elevated ovarian cyp19a1a expression, and there was a significant interaction with inbreeding (P = 0.0147) according to the best-fit lme model (SI Appendix, Table S1.5).