Figure 5.
Expression of Ad4BP/SF-1 in the M33-KO spleens. To investigate the expression levels of Ad4BP/SF-1, Western blot analysis was performed using total cell lysates from the spleens of wild-type (+/+), heterozygous (+/–), and homozygous (–/–) knock-outs at 16.5 days after coitus (A). As the control, α-tubulin expression was examined. The relative levels of Ad4BP/SF-1 expression were quantified (B). Data represent the means ± SD of 3 independent pools of spleens. The amount of Ad4BP/SF1 mRNA was analyzed by RT-PCR using total RNA from the spleens of wild-type (+/+), M33 heterozygous (+/–), and homozygous (–/–) knock-outs at 16.5 days after coitus (C, top). GAPDH was used as a control (C, bottom). Immunohistochemical experiments were performed on wild-type (D, right) and M33-KO spleens (D, left) using an antibody against Ad4BP/SF-1 at 16.5 days after coitus. Closed arrowheads (D) indicate Ad4BP/SF-1–immunoreactive cells lining the vascular tubules, whereas open arrowheads indicate the vascular tubules in the M33-KO spleen. The expression of M33 was analyzed immunohistochemically using the antibody to M33 in the wild-type (E, right) and Ad4BP/SF1-KO spleens (E, left) at 16.5 days after coitus. Closed arrowheads (E) indicate M33 immunoreactive cells lining at the vascular tubules. Scale bar, 20 μm.