Fig. 5.

Transient transgenic analysis of a DHS unique to Sertoli cells identifies a new Sertoli cell enhancer upstream of Wt1. (A) DNaseI-seq and H3K27ac ChIP-Seq data around the Wt1 locus. Gene names indicate the TSS of the transcript. Nearby genes are indicated in gray. DNaseI-Seq data (Parzen score) and peak calls (boxes above track) are in light blue; H3K27ac ChIP-seq data and peak calls are in dark blue. A box surrounds the DHS chosen for in vivo analysis. (B) Detailed view of the DHS showing the DNaseI-seq Parzen score, the smoothed base counts (light blue) and the H3K27ac data (dark blue). The light- and dark-blue bars indicate the DNaseI-seq and H3K27ac peaks, respectively. The black bar marks the region cloned upstream of an hsp68-LacZ reporter cassette. (C) An E13.5 testis from a transgenic embryo showed β-galactosidase expression (TgWt1, green) specifically in Sertoli cells. Sertoli cells were labeled by AMH immunostaining (red) and germ cells by CDH1 (blue). The confocal image was taken using a 20× objective. Scale bar: 100 µm. (D) Confocal image of the sample in C taken with a 40× objective. Scale bar: 12.5 µm. All three panels show germ cells (blue); the left panel shows Sertoli cells (red); the middle panel shows β-galactosidase expression (green); the right panel shows the merge (yellow indicates co-expression of β-galactosidase and AMH).