Figure 1. Sf-1^2KR/2KR mice exhibit disrupted endocrine physiology without affecting SF-1 protein levels.

A. Protein lysates from Sf-1^+/+ E16.5 gonads were immunoprecipitated (IP) and western blotted (WB) using anti-SF-1 antibodies. Mono- (1xSu-SF-1) and di- (2xSu-SF-1) sumoylated SF-1 are indicated (arrowheads) with a lighter exposure of unmodified SF-1 shown in bottom panel. B. The targeted allele of unsumoylatable SF-1 mutant mouse (Sf-1^2KR/2KR) with individual mutations shown in Exon 3, and before removal of Neo cassette (Neo). C. Sumoylation of SF-1 is abolished in Sf-1^2KR/2KR mice. SF-1 levels are shown for postnatal day (P) 3 wild type and mutant tissues with βactin as loading control. D. Western blots of E16.5 ovaries and testes (left panels), and qPCR of E16.5 testes and adrenals (right panels) showed no difference in SF-1 protein and transcript levels, respectively. E. Immunostaining of SF-1 in wild type and mutant E16.5 adrenals. F. Testicular/adrenal weights and sperm counts are shown for 8 or 12 weeks old Sf-1^2KR/2KR males, respectively, n=7 for Sf-1^+/+ and n=6 for Sf-1^2KR/2KR. G. Gonadal testosterone (T) levels and adrenal corticosterone (Cort) levels measured in P23 male wild type and Sf-1^2KR/2KR littermates. All data points in steroid assays passed the Grubbs’ outliers test. See also Figure S1, S2 and Table S1.