Figure 4.

Phenotypic and gene expression changes in E12.5 XY Sox9; Wnt4 mutants (A–D). Immunodetection of the pre-granulosa cell marker FOXL2 (green) and the male supporting cell marker SRY (red) in E12.5 XY gonads of Control (A), Sox9^cKO (B), Wnt4^KO (C), and Sox9^cKO Wnt4^KO (D) genotypes. (E–H) Immunodetection of the Sertoli cell marker AMH (green) in E12.5 XY gonads of Control (E), Sox9^cKO (F), Wnt4^KO (G), and Sox9^cKO Wnt4^KO (H) genotypes. (I–L) Immunodetection of the steroidogenic enzyme HSD3B (red) in E12.5 XY gonads of Control (I), Sox9^cKO (J), Wnt4^KO (K), and Sox9^cKO Wnt4^KO (L) genotypes. (M–R) RT-quantitative PCR analysis of Foxl2 (M), Axin2 (N), Follistatin (Fst) (O), Amh (P), Sox8 (Q), and Fgf9 (R) expression in E12.5 gonads of XX Control, XY Control, XY Sox9^cKO, XY Wnt4^KO, and XY Sox9^cKO Wnt4^KO genotypes (N = 3–4 embryos for each genotype). Expression level in XX controls is 1. Graphs show individual values (dots), and the mean fold-change (bars) ± SEM. Nuclei labeled with DAPI are shown in blue. Magnification is the same in all panels. Scale bar = 100 μm. Gonads are outlined with broken white lines. The white arrow in D indicates the coelomic vessel.