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p53 MO Attenuates Cell Death Induced by MOs and GripNAs (A–G and I–N) Brightfield images of 28 hpf embryos injected with demonstrative MOs and gripNAs. The arrows (C, E, I, and M) indicate neural death that is significantly attenuated to normal head size and morphology by co-knockdown of p53 (D, F, J, and N). Interestingly, Wnt5 MO2 shows no significant neural death (G) even at 6 ng, a higher dose than Wnt5 MO1 (E) (3 ng), but can elicit a highly penetrant Wnt5 phenotype even at 1.5 ng (unpublished data). (H) Effect of p53 MO on Wnt5 splicing. We carried out RT-PCR using primers spanning the exon 5–exon 6 junction targeted by Wnt5 MO1. Proper splicing was completely inhibited by Wnt5 MO1. p53 co-knockdown did not affect the efficiency of Wnt5 MO1 inhibition. Embryos injected with Wnt5 MO2, which targets the previous junction (exon 4–exon 5), still exhibited some properly spliced transcript at the exon 5–exon 6 junction. β-actin was used as a loading control.
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