Xenbase Image ID: 48876

Fig. 9. Vent and Sia directly act on the hhex promoter. (A) Animal cap explants injected with GR-Vent2–VP16 RNA (100 pg) were cultured with or without CHX for 30 min and then treated with DEX. In situ hybridization at stage 11 shows that hhex and otx2, but not chordin, were directly induced by GR-Vent2–VP16 in CHX-treated explants. (B) The indicated hhex:luc reporters were injected into either C1 dorsal (green) or C4 ventral cells (red), with either Vent1/2-MOs (30 ng each) or Sia mRNA (50 pg). The histogram shows the average normalized luciferase activity at stage 10.5 and standard deviation from a single representative injection experiment performed in biological triplicate. *p < 0.05 in Student T-test when compared to injection of reporter alone and ns = no statistical difference. A representative example of 3 independent injection experiments is shown. (C) GR-Sia can directly activate hhex transcription in Vent-depleted animal caps. Animal caps injected with GR-sia RNA (30 pg) with or without Vent1/2-MOs (30 ng each) were cultured with CHX, DEX, or CHX + DEX followed by in situ hybridization at stage 11 for hhex or chordin mRNA. (A, C) Animal cap experiments were repeated twice with identical results and a total of ~ 10 explants/condition. One experiment is shown. Image published in: Rankin SA et al. (2011) Copyright © 2011. Image reproduced with permission of the Publisher, Elsevier B. V. Experiment + Assay Source Phenotypes and Disease Xla Wt + VP16-ventx2.2-GR + CHX + animal cap explant + NF11 (in situ hybridization) fig.9.a Expression Phenotype decreased amount chrd.1.L expression in animal cap decreased amount hhex.L expression in animal cap decreased amount otx2.L expression in animal cap Xla Wt + VP16-ventx2.2-GR + CHX + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.a Expression Phenotype decreased amount chrd.1.L expression in animal cap increased amount hhex.L expression in animal cap increased amount otx2.L expression in animal cap Xla Wt + VP16-ventx2.2-GR + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.a Expression Phenotype increased amount chrd.1.L expression in animal cap increased amount hhex.L expression in animal cap increased amount otx2.L expression in animal cap Xla Wt + sia1-GR + CHX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype decreased amount chrd.1.L expression in animal cap decreased amount hhex.L expression in animal cap Xla Wt + sia1-GR + CHX + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype increased amount chrd.1.L expression in animal cap decreased amount hhex.L expression in animal cap Xla Wt + sia1-GR + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype increased amount chrd.1.L expression in animal cap increased amount hhex.L expression in animal cap Xla Wt + sia1-GR + ventx1.1 MO + CHX + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype increased amount chrd.1.L expression in animal cap increased amount hhex.L expression in animal cap Xla Wt + sia1-GR + ventx1.1 MO + ventx2.1 MO + CHX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype decreased amount chrd.1.L expression in animal cap decreased amount hhex.L expression in animal cap Xla Wt + sia1-GR + ventx1.1 MO + ventx2.1 MO + DEX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype increased amount chrd.1.L expression in animal cap increased amount hhex.L expression in animal cap Xla Wt + ventx1.1 MO + ventx2.1 MO + CHX + animal cap explant + NF11 (in situ hybridization) fig.9.c Expression Phenotype decreased amount chrd.1.L expression in animal cap decreased amount hhex.L expression in animal cap Larger Image Printer Friendly View

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