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Fig. 5. Microinjection of Xfoxi1a mRNA induces epidermal differentiation and suppresses neural induction in both in vivo and in vitro. Xfoxi1a mRNA (25 pg/cell) was injected into two unilateral blastomeres of eight-cell embryos. Embryos were fixed at stage 14-15 and then whole-mount in situ hybridization was performed with the following probes. (A) Sox2, (B) XK81, (C) Dlx3, (D) FoxD3 and (E) Six1. (A-D) Dorsal view (anterior towards top); (E) anterior view (dorsal towards the top). Dashes indicate the midline. White arrows indicate the injected side. The activity of Xfoxi1a (12.5 pg/cell) was assessed by RT-PCRs in Chd (50 pg/cell)-injected animal caps (F) or dominant-negative Bmp receptor (dnBMPR) (100 pg/cell)-injected animal cap (G). RNAs were injected into all the animal blastomeres of eight-cell embryos. The animal caps were excised at stage 9 and cultured until sibling embryos reached stage 15. The expression patterns of the neural marker Sox2, the non-neural ectodermal markers (XK81, Dlx3, Msx1, Xfoxi1a, Bmp4), the mesodermal marker MyoD were analyzed. H4 was used as the loading control. |