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Fig. 3. Depletion of zygotic NF2 reduces anterior development. (A) Depiction of a part of nf2 cDNA sequence spanning the start codon (boxed) and the sequences complementary to the NF2 MOs. (B) Western blotting analysis for the endogenous NF2 protein in the control MO (cMO) and NF2 MO1-, or MO2-injected embryos at different developmental stages. β-Actin was used as a loading control. Each MO was injected at the dose of 40 ng per embryo. The numerical numbers indicate the relative expression levels of NF2 protein after normalized to the levels of β-actin. (C) Early tadpole stage embryos (st39/40) showing the phenotypic effects of zygotic NF2 depletion. Red arrows point to the encircling eyes. (D) Early tailbud stage (st23/24) embryos in situ hybridized with six3 and hoxb9. Upper panels: anterior view. Lower panels: dorsal view with anterior upwards. (E, F) Early tailbud stage embryos (dorsal anterior view) in situ hybridized with krox20, en2, and rx. (G) Western blotting analysis of NF2 protein in embryos at stage 15 with the indicated treatments. NF2 MOs were injected into both cells at 2-cell stage; MO-insensitive nf2 mRNA (200 pg) was injected into all four cells at 4-cell stage. The resultant embryos were lysed at stage 15. β-actin was used a loading control. |
