| |
Figure 3. Overexpression of δNp63 induces epidermal fate and suppresses neural induction. A–F: δNp63 mRNA (700 pg) was injected into one dorsal blastomere of 16-cell stage embryos. Embryos were fixed at stage 16 and whole-mount in situ hybridization was performed for (A) XK81a, (B) Dlx3, (C) Rexp52, (D) XAG, (E) FoxD3, and (F) Sox2. A–C,E,F: Dorsal view, anterior on the right. D: Anterior view, dorsal at the top. Dashed lines indicate the midline, small arrowheads indicate increased expression, and brackets indicate the width of the neural plate. Arrowheads indicate the injected side, recognized by FLDx staining. G–L: One-cell stage embryos were injected with 500 pg of Chd (H,K) or 500 pg of Chd + 500 pg δNp63 mRNA (I,L). G,J: Control embryos were not injected. Embryos were cultured until the blastula stage when animal caps were dissected out and processed when sibling embryos reached the equivalent of stage 16. The expression of XK81a and Sox2 was assessed by in situ hybridization. M: Total RNA was isolated from treated and control caps and the expression of δNp63, epidermal markers XK81a and Dlx3, neural marker Sox2 and the mesodermal marker XBra was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR).Download figure to PowerPoint |
