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Fig. 5. March2 is required for anterior head formation in Xenopus. (A) Dorso-animal injection of Mar2MO (40 ng) at the eight-cell stage results in defects in anterior head formation; these defects were rescued by co-expression of a MO-resistant form of Xenopus March2 mRNA (500 pg). (B) Quantification of A. (C) In situ hybridization analysis showing that Mar2MO (40 ng) suppresses expression of Chd at stage 12, and Gsc and Otx2 at stage 11.5. Co-expression of March2 mRNA (500 pg) rescues the reduction of expression of these genes (see Fig. S5A). (D) Mar2MO suppresses expression of BF1, Otx2, En2 and Krox20 at stage 17. CoMO (40 ng) or Mar2MO (40 ng) along with β-galactosidase mRNA (100 pg) as a lineage tracer were injected into one dorso-animal blastomere of eight-cell stage embryos. The right side of the embryos shown have been injected with indicated reagents (see Fig. S5B). (E,F) March2 regulates head formation through Dsh. Two dorsoanimal blastomeres of eight-cell stage embryos were injected with indicated reagents, and phenotypes were analyzed at tadpole stages. (E) Mar2MO (20 ng) along with Dsh mRNA (250 pg) synergistically induced head defects. (F) Head defects caused by Mar2MO (60 ng) were partially rescued by DshMO (40 ng). Statistical analysis was performed on three independent experiments. P values were obtained using two-tailed Student’s t-test. For the number of embryos in each quantification, see Table S3. |
