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. 2011 Sep 7;6(9):e24469. doi: 10.1371/journal.pone.0024469

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Xie et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) The foxo3b-ATG-MO injected embryos showed remarkable loss of expression of anterior neural markers by 24 hpf, which could partially be rescued by co-injection of foxo3b mismatch mRNA. (A1–A3, A13) The expression of nkx5.1 at the telencephalon (indicated by white arrows) was dramatically reduced in foxo3b-ATG-MO injected embryos. Co-injection of foxo3b mismatch mRNA could partially restore its expression at the telencephalon. (A4-A6, A14) Six3b was specifically expressed at the telencephalon and eyes. Loss of foxo3b function resulted in abnormal expression pattern of six3b, which was restored by co-injection of foxo3b mismatch mRNA. (A7–A9, A15) Pax6 expression at the forebrain and eyes decreased greatly in foxo3b morphants. Co-injection of foxo3b mismatch mRNA partially restored its expression. (A10–A12, A16) Expression of tbx5 at the retina was dramatically reduced in foxo3b-knockdown embryos. Its expression was rescued by co-injection of foxo3b mismatch mRNA. (A17) Opl expression at the telencephalon was reduced in foxo3b morphants compared to control embryos, which was efficiently rescued by co-injection of foxo3b mismatch mRNA. Embryos were injected with 8 ng foxo3b-ATG-MO or 125 pg foxo3b mismatch mRNA,wild-type embryos were used as control. A1-A3, lateral views with anterior to the left; A4-A12, dorsal views with anterior to the left; A1-A17, 24 hpf. (B) The foxo3b-SP-MO injected embryos exhibited anterior defects similar to that of foxo3b-ATG-MO injected embryos. (B1–B2, B7) The expression of pax6 at the telencephalon and eyes was reduced in foxo3b-SP-MO injected embryos. (B3–B4, B8) Tbx5 expression at the eyes decreased in foxo3b-knockdown embryos. (B5–B6, B9) Loss of zygotic foxo3b function resulted in reduction of nkx5.1 expression at the telencephalon. Embryos were injected with 16 ng STD-MO (control) or 16 ng foxo3b-splice-MO. B1–B4, dorsal views with anterior to the left; B5-B6, lateral views with anterior to the left; B1-B9, 24 hpf.