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Fig. 5. Prdm13 is activated by the Ptf1-J transcription complex. (A) In situ hybridization analysis of Prdm13 and Gad1 in embryos injected with Ptf1a-GR, Ptf1a/bHLH-Neurog2-GR or Ptf1aW224A/W242A-GR. Dorsal views, anterior to the bottom, of neurula embryos are shown for Prdm13. Whole head dorsal views and transverse sections at the level of the posterior hindbrain are shown for Gad1. (B) Real time RT-PCR analysis of the expression of Prdm13 in stage 16 animal cap explants isolated from embryos overexpressing Ptf1a-GR, Ptf1a/bHLH-Neurog2-GR or Ptf1aW224A/W242A-GR. Note that Prdm13 is dramatically activated in both assays by Ptf1a and to a lesser extent by Ptf1a/bHLH-Neurog2-GR but not by Ptf1aW224A/W242A-GR, which is also unable to activate Gad1. (C) In situ hybridization analysis of Prdm13 and Neurog2 in embryos injected with Ptf1a, Rbp-J, or a combination of Ptf1a and Rbp-J. Dorsal views, anterior to the bottom, of neurula embryos are shown. (D) Real time RT-PCR analysis of the expression of Prdm13 and Neurog2 in stage 16 animal cap explants isolated from embryos overexpressing Ptf1a, Rbp-J, or a combination of Ptf1a and Rbp-J. Note that Rbp-J does not affect Prdm13 nor Neurog2 expression and that the combination of Ptf1a and Rbp-J induces stronger Prdm13 and Neurog2 expression than that induced by Ptf1a alone. In all cases, LacZ mRNA was used as a lineage tracer to identify the injected side. IS, injected side. Image published in: Hanotel J et al. (2014) Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V. Experiment + Assay Source Phenotypes and Disease Xla Wt + ptf1a-GR + DEX + NF15-16 (in situ hybridization) A, H Expression Phenotype increased amount prdm13.L expression in neural plate Xla Wt + ptf1a-GR + DEX + tailbud stage (in situ hybridization) A, H Expression Phenotype increased amount gad1.1.L expression in neural tube Xla Wt + ptf1a-neurog2-GR + DEX + NF15-16 (in situ hybridization) A, H Expression Phenotype mislocalised prdm13.L expression in neural plate increased amount prdm13.L expression in neural plate Xla Wt + ptf1a-neurog2-GR + DEX + tailbud stage (in situ hybridization) A, H Expression Phenotype increased amount gad1.1.L expression in neural tube Xla Wt + ptf1a + neurula stage (in situ hybridization) C, D Expression Phenotype increased amount neurog2.L expression in neural tube mislocalised prdm13.L expression in ectoderm increased amount prdm13.L expression in ectoderm mislocalised prdm13.L expression in neural plate increased amount prdm13.L expression in neural plate Xla Wt + ptf1a + rbpj + neurula stage (in situ hybridization) C, D Expression Phenotype mislocalised neurog2.L expression in neural tube increased amount neurog2.L expression in neural tube mislocalised prdm13.L expression in neural plate increased amount prdm13.L expression in neural plate Larger Image Printer Friendly View

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