FIG. 3.

Sertoli cell-specific expression of Cbln4. A) Real-time RT-PCR analysis of wild-type (wt) and homozygous W^e/W^e littermate gonads for Cbln4, Sox9 (Sertoli cell marker), and Mvh (germ cell marker) relative to 18S RNA (mean ± SEM of three independent experiments). Individual experiments were performed in triplicate on RNA obtained from pooled gonads from three to four littermates. Cbln4 showed expression similar to Sox9 and is maintained in the germ cell-depleted gonads. In comparison, Mvh expression was lost in W^e/W^e samples. B) In situ hybridization on sagittal paraffin sections showed Cbln4 (right column) and Amh (left column) expression specifically in Sertoli cells at 13.5 dpc, whereas the germ cell marker Pou5f1 was expressed by germ cells only (middle column). The bottom row shows enlargements (magnified ×4) of the regions marked with rectangles in the top row. Images are oriented so that the anterior pole is at the top. Bar = 200 μm.