Fig. 7

Maternal Vsx1 directly represses ntl ectopic expression. (A-I) Expression of ntl in uninjected wild-type (A-C), vsx1 tbMO-injected (D-F) and vsx1 mRNA-injected (G-I) embryos at 30% epiboldy. (B, E, H) Dorsal views with animal towards the top. (C, F, I) Animal views with dorsal towards the right. (J) Real-time quantitative RT-PCR analysis of ntl expression in vsx1 knockdown or overexpression embryos compared to uninjected wild-type embryos. Results are expressed as mean±SEM, and statistical analyses were done by unpaired t test. **P<0.01. ***P<0.001. (K) The position of potential Vsx1 binding sites at ntl proximal promoter region. B1-B18 indicates the potential binding sites of Vsx1. R1-R8 represents the examined regions in ChIP assay. (L) ChIP assay on extracts from wild-type embryos. Input is a positive control with sonicated original genomic DNA fragment. No antibody immunoprecipitates are used as negative controls for ChIP assay specificity. These results show the specific recruitment of Vsx1 by ntl promoter region 8. (M) Diagram of GFP reporters driven by wild-type or mutant fragments of the ntl proximal promoter. (N) GFP expression driven by wild-type or mutant fragments of the ntl proximal promoter. The types of GFP reporter sensors are shown at the top of the images and the coinjected mRNA is shown at the left of the images.