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. 2009 Feb 9;29(8):2168–2180. doi: 10.1128/MCB.01508-08

FIG. 5.

FIG. 5.

Overexpression of Myt1 results in apoptosis during gastrulation. (A) Xenopus embryos were injected at the one-cell stage with the mRNA of Myt1 at the indicated dose or 100 pg of β-Gal (negative control). Embryos were collected, and lysates were subjected to immunoblotting using anti-Flag. (B) Xenopus embryos injected with the mRNA of β-Gal or human Myt1 were photographed. Control, normal embryos at stage 12 derived from one-cell-stage embryos that were injected with the mRNA of β-Gal (100 pg) as a control; Type A, Type B, and Type C, embryos derived from one-cell-stage embryos that were injected with the mRNA of Myt1. The types of embryos were classified as type A, type B, or type C based on the severity of the phenotypic defect. (C) Histogram representing the number of phenotypic changes induced by Myt1. The asterisks indicate a significant increase in lethality compared to that for the control (P < 0.05). These data are representative of the results from three independent experiments. (D) Whole-mount TUNEL assay. Albino embryos injected with the mRNA of Myt1 (50 pg) or β-Gal (50 pg) were fixed at stage 11 and processed for TUNEL staining. A positive TUNEL reaction is indicated by purple staining of the nuclei. (E) One-cell-stage Xenopus embryos were injected with the mRNA of Myt1 (70 pg) with/without that of DN-JNK1 (0.5 or 1.0 ng). The histogram represents the number of phenotypic changes. The asterisks indicate a significant decrease in dissociation compared to that for the control (P < 0.05). These data are representative of the results from three independent experiments. (F) One-cell-stage Xenopus embryos were injected with the mRNA of Myt1 (50 pg) with/without human JNK1 or human JNK2 (200 pg of each). The embryos shown were photographed at stage 12. (G and H) Histograms representing the number of phenotypic changes induced by Myt1 with/without JNK1 (G) or JNK2 (H). The asterisks indicate a significant increase (G) or decrease (H) in dissociation/death compared to that of the control (P < 0.05). These data are representative of the results from three independent experiments.