XB-IMG-23575
Xenbase Image ID: 23575
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Figure 4. XFDL Antagonizes the Activation of Mesodermal Marker Genes by p53(A–F) In situ hybridization analysis of early gastrulae. Side view. Control mRNA (250 pg/cell; A and D), human p53 (hp53; 50 pg/cell; B and E), or hp53 + 400 pg/cell of XFDL (C and F) mRNAs were injected radially at the 4-cell stage.(G) q-PCR analysis for Mix.2 expression. Control (lane 1), hp53 (lanes 2–5), hp300 (lanes 3–5), 100 pg/cell of XFDL (lane 4), and/or 400 pg/cell of XFDL (lane 5) mRNAs were injected. Animal caps were prepared at stage 8.5 and harvested at stage 11.(H–L) Double knockdown analysis of XFDL and p53. (H–K) Embryos injected with XFDL-MO (50 ng/cell; I and K) and/or p53-MO (50 ng/cell; J and K) were harvested at stage 10.5 and analyzed by in situ hybridization with the Xbra probe. (L) q-PCR analysis using the animal cap. XFDL-MO (50 ng/cell; lanes 2–4 in each panel) with p53-MO (12.5 ng/cell for lane 3 and 50 ng/cell for lane 4 in each panel) was injected and analyzed as in (G).(M) The reporter constructs used in (N) to (S).(N–T) Luciferase assays in the animal caps injected with hp53, hp300, and XFDL mRNAs. mRNAs (the same as in G) were coinjected with the indicated reporter plasmids (50 pg/cell each). Animal caps were prepared as in (G) and dual-luciferase assays were performed. In (N), (P), (R), and (T), caps were treated with Activin (5 ng/ml). Error bars represent the standard deviation (SD).(U) q-PCR analysis for Mix.2 expression. GR-hp53 mRNA (50 pg/cell; lanes 2, 3, 5, and 6) was injected without (blue columns) or with (red columns) XFDL-MO (50 ng/cell), and animal caps were prepared at the mid-gastrula stage (stage 8.5). Dexamethazone (Dex; 10 μg/ml) was performed from stage 8.5 to stage 10 (lane 3) or from stage 11 to stage 12 (lane 6). Image published in: Sasai N et al. (2008) Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V.
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