Figure 2From: Disruption of genital ridge development causes aberrant primordial germ cell proliferation but does not affect their directional migrationWt1 is essential for epithelial cells characteristics maintenance and their normal proliferation. TUNEL assay of coelomic epitheliums (white lines) was detected in Wt1R394W/R394W embryo and control embryo (A). A table outlining the number of TUNEL+ PGCs observed in control and Wt1R394W/R394W E11.5 embryos (C). Proliferation of coelomic epitheliums (white lines) in Wt1R394W/R394W embryos was examined, using BrdU incorporation assay (B). White arrows in B pointed to the BrdU+ coelomic epitheliums. Scale bars: 30 μm. Numbers in parentheses show the actual number of BrdU+ coelomic epitheliums over the total number of coelomic epitheliums counted for that embryo (D), and note that the BrdU labeling rate was significantly reduced in Wt1R394W/R394W embryos (E). Four potential WT1 binding sites (GNGGGNG) in the cyclin E1 2.8-kb promoter were identified (F). TM4 cells were co-transfected with 0.2 μg cyclin E1-Luc (G), or 0.2 μg cyclin E1-Luc with different point mutants of the WT1A in modified PCB6+ vector (H), or the control PCB6+ vector. One group of mutants contained changes in the zinc finger region (R366C, H337Y and R394W), and the other group contained (F154S and S273A) changes outside that zinc finger region [37]. All of these assays were performed 36 hours after transfection. Fold activation was displayed by Firefly/Renilla ratio. Error bars represent SEM. *P <0.05 (by t-test). A significant elevation of mesenchymal markers vimentin (left) and a complete loss of the epithelial markers E-cadherin (right) and were detected by immunostaining assay (I). White arrows in I pointed to the signals. Scale bars: 30 μm.Back to article page