PGC proliferation was decreased in GR-deficient embryos. Cell apoptosis and proliferation were assessed by TUNEL (A-F, green) and BrdU incorporation (G-L, green) assays, respectively. The PGCs were labeled with Stella (red). Table outlining the number of TUNEL-positive PGCs observed in control and Wt1R394W/R394W embryos (M). Note that no TUNEL-positive PGCs were observed in either control (C) or Wt1R394W/R394W (F) embryos at E11.5. Percentage of total PGCs that are BrdU+. Numbers in parentheses show the actual number of BrdU+ PGCs (Stella+ BrdU+ cells, white arrows) over the total number of PGCs counted for that embryo (Stella+ cells) (N). Note that approximately 50% of the PGCs were labeled with BrdU (I, white arrows, O) in control embryos. In contrast, only approximately 28% of the PGCs in Wt1R394W/R394W embryos were labeled with BrdU (L, white arrows, O). Scale bars: 30 μm. The mRNA level of BMP family members (Bmp2, Bmp4, Bmp5 and Bmp6), TGF-β family members (Tgfβ1 and Activin), and downstream genes of BMP signaling (Smad3, Smad4, Smad5 and Smad8) was detected by real-time RT-PCR in male (P) and female (Q). The mRNA level was normalized against Gapdh.* P <0.05. Each bar, mean ± SD of n = 3 experiments.