irx4a morphant embryos show defects in head size and extensive cell death. (A, B) Images of live wild type (A) and morphant (B) embryos at 24 hpf; Compare the size of the eyes and head (arrows). (C, D) Acridine orange staining of wild type (C) and irx4a morphant (D) embryos viewed at 24 hpf. Note the increased number of labeled cells in the morphant embryo (E, F) MO4as injected embryos do not show substantial changes in cell proliferation as evidenced by anti-phospho-histone H3 (H3-P) immunostaining. (G) RT-PCR showing the effect of the MO4as on irx4a transcript splicing. RNA prepared from uninjected embryos (“C”) shows the internal control band (actin, act) but the absence of unprocessed irx4a mRNA. The presence of this band in MO4as injected fish (arrow) indicates defective irx4a mRNA splicing. Panels A–F show lateral views of 24 hpf embryos; dorsal is up, anterior is left. Scale bar in F: 70 μm.
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