The Sox9 and Cyp19a1 expression change in response to Amh overexpression. (A and B) qRT-PCR of Sox9 and Cyp19a1 in control ZW, ZW + Amh-OE, and control ZZ gonads of stage 25, showing increased Sox9 expression and reduced Cyp19a1 expression in ZW gonads overexpressing Amh. Data are shown as means ± SD. N ≥ 3. ** P < 0.01; *** P < 0.001. (C) Double immunofluorescence of SOX9 and CTNNB1 in sections of control ZW, ZW + Amh-OE, and control ZZ gonads of stage 25. SOX9 protein was induced to express robustly in gonadal medulla of ZW embryos overexpressing Amh. Bar, 50 μm.