Adrenal defects in the homozygous M33- and heterozygous Ad4BP/SF1-KO mice. (A) Macroscopic comparison of adrenal glands harvested at 18.5 days after coitus from M33-KO (M33^–/–), wild-type (M33^+/+), and Ad4BP/SF1 heterozygous KO (Ad4BP/SF-1^+/–). Scale bar, 400 μm. Stereomicroscopic images were photographed and processed as described in Figure 1A-D. (B) To investigate the expression levels of Ad4BP/SF-1, Western blot analysis was performed using total-cell lysates from the adrenal glands of wild-type (^+/+), heterozygous (^+/–), and homozygous (^–/–) M33 KO at 16.5 days after coitus. α-Tubulin expression was examined as the control. The relative levels of Ad4BP/SF-1 expression were quantified by National Institutes of Health (NIH; Bethesda, MD) image analysis software (C). The amount of Ad4BP/SF-1 mRNA was analyzed by quantitative RT-PCR (Roche) using total RNA from the adrenal glands of wild-type (^+/+), M33 heterozygous (^+/–), and homozygous (^–/–) KO at 16.5 days after coitus (D). (C,D) Data represent the mean ± SD values of 3 independent pools of the adrenal glands.