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Experiment details for th

Wylie LA et al. (2015) Assay

Ascl1 phospho-status regulates neuronal differentiation in a Xenopus developmental model of neuroblastoma.

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Gene Clone Species Stages Anatomy
th.S laevis NF stage 18 to NF stage 23 anterior placodal area

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  Fig. 2. Ascl1 is expressed transiently during noradrenergic development in the anteroventral region Xenopus embryos were fertilized and allowed to develop to the developmental stage indicated before fixing and staining by ISH for NA neuron markers Ascl1, Hand2, Phox2a, and TH along with the neural crest marker Sox10. Embryos are all orientated with the ventral side imaged and head to the top. The anteroventral region where AVNA cell markers are expressed is expanded in the lower panel.

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Gene Clone Species Stages Anatomy
th.S laevis NF stage 18 to NF stage 19 anterior placodal area

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  Fig. 3. p27Xic knock-down by morpholino injection inhibits neurogenesis Xenopus laevis embryos were injected in one of the two-cell stage with beta-galactosidase (light blue) as a tracer along with either Xic1 mRNA or Xic1 antisense morpholino (MO) as indicated, injected side to the right. (A) At stage 19-20 embryos were subject to in situ hybridization to detect neural-β-tubulin, Phox2a, Hand2 and TH. (B) Embryos were scored for marker expression on the injected side relative to the uninjected side. For scoring data, [n=12-34]. Kruskal-Wallis non-parametric ANOVA was performed on control MO compared to Xic1 MO and uninjected compared to Xic1 mRNA (p-values of <0.05 = *, <0.0005 = **, and <0.000005 = ***).

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Gene Clone Species Stages Anatomy
th.S laevis NF stage 18 to NF stage 19 anterior placodal area

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  Fig. 6. CDK activity inhibits AVNA cell differentiation induced by WT Ascl1 but not SA Ascl1 Xenopus embryos were injected in one of the two-cell stage with beta-galactosidase (light blue) as a tracer along with mRNA encoding Cyclin A/CDK2 and/or mouse WT/S-A Ascl1, as indicated, injected side to the right. (A) Embryos were subject to in situ hybridization for AVNA markers at stage 18-19, as labeled, (see Supplementary Fig. 2 for scoring scheme). (B) Embryos were scored for marker expression comparing the injected and ininjected side. For scoring data, [n=20-32]. Kruskal-Wallis non-parametric ANOVA was performed comparing WT Ascl1 with S-A Ascl1 without and with Cyclin A/CDK2 (p-values of <0.05 = *, <0.0005 = **, and <0.000005 = ***).

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Gene Clone Species Stages Anatomy
th.S laevis NF stage 18 to NF stage 19 anterior placodal area

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  Fig. 7. N-Myc inhibits AVNA cell differentiation induced by WT Ascl1 but not S-A Ascl1 Xenopus embryos were injected in one of the two-cell stage with beta-galactosidase (light blue) as a tracer along with mRNA encoding N-Myc with or without mouse WT/S-A Ascl1, as indicated. (A) Embryos were subject to in situ hybridization for AVNA markers as indicated at stage 18-19. (B) Embryos were scored for marker expression comparing the injected and ininjected side. For scoring data, [n=24-32]. Kruskal-Wallis non-parametric ANOVA was performed comparing WT Ascl1 and S-A Ascl1 with and without N-Myc (pvalues of <0.05 = *, <0.0005 = **, and <0.000005 = ***).