DEV00957 Supplemental Data

Files in this Data Supplement:

• Figure 1 -

  Fig. S1. Characterization of Ar exon 1 inversion allele. (A) Cre-recombinase activity in Ar^{flox(ex1-neo)/Y} males causes complete androgen insenitivity. Ar^{inv(ex1-neo)/Y}; Sycp1-cre animals are female in external appearance. Blue arrowheads indicate presence of abdominally positioned testes. All other internal reproductive organs are absent. (B) RT-PCR analysis of total testis RNA. Wild-type Ar RNA is absent from Ar^{inv(ex1-neo)/Y} males, while an exon1-neomycin fusion RNA is generated from the inverted allele.

• Figure 2 -

  Fig. S2. Generation of Amh-cre transgenic animals. (A) Cre cDNA inserted into Amh exon 1 fragment. Known Amh promoter elements are shown. Cre cDNA is followed by full length Amh intron 1. (B) Assay for b-galactosidase activity. CRE function is limited to testes of e14.5Amh-cre fetuses. No staining is detected in attached

• Figure 3 -

  Fig. S3. Characterization of testicular AR protein expression. (A,B) AR protein is detected in Sertoli cells of Ar^{flox(ex1-neo)/Y}; Amh-cre and Ar^+/Y testes by an N-terminal directed antibody [AR (N)]. Red arrowheads indicate AR-positive Sertoli cells. (C) A C-terminal specific antibdy [AR(C-19), Santa Cruz Biotechnology] fails to detect AR in Sertoli cells of Ar^{flox(ex1-neo)/Y}; Amh-cre testes, demonstrating the absence of full-length AR. Black arrowheads indicate AR-negative Sertoli cells. (D) Full-length AR protein is detected in wild-type testes by the C-terminal-specific antibody.