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Gene/Clone	Species	Stage	Anatomy Item	Experimenter
tbxt	xenopus

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Experiment details for tbxt

Sasai N et al. (2008) Assay

Ectodermal factor restricts mesoderm differentiation by inhibiting p53.

Gene	Clone	Species	Stages	Anatomy
tbxt.S		laevis	NF stage 10	mesoderm

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Figure 3. Mesodermal Conversion Induced by XFDL Knockdown in the Animal Cap(A–J) In vivo effects of XFDL-MO. Control embryos (A, E, G, and I) or embryos given 50 ng/cell of XFDL-MO (25 ng/cell of XFDL156-MO + 25 ng/cell of XFDL141-MO for the short minor splicing variant; see Supplemental Experimental Procedures; B, F, H, and J), two five-base-mismatched MOs (25 ng/cell of each; C), or 50 ng/cell of XFDL-MO + XFDL mRNA (100 pg/cell; D) by injection were harvested at the early gastrula stage and analyzed by in situ hybridization using the indicated probes. Sectioned embryos are shown in (E) and (F). Brackets (in panels A and B) and arrowheads (in panels E and F) indicate the expression areas of Xbra and VegT, respectively.(K) q-PCR analysis for the expression of mesodermal marker genes. XFDL-MO (50 ng/cell) was injected at the 8-cell stage. Animal caps were prepared at stage 8.5, cultured until siblings reached stage 10.5, and analyzed by q-PCR.(L) Chordin-induced neural differentiation was attenuated by XFDL-MO in the animal cap. Control (10 pg/cell; lane 1) or Chordin (10 pg/cell; lanes 2–4) mRNAs were injected with 12.5 ng/cell (lane 3) or 50 ng/cell (lane 4) of XFDL-MO. Explants were prepared at stage 8.5 and harvested at stage 11.5. Sox2 expression was analyzed by q-PCR. In (K) and (L), the expression of each gene in the control animal cap was defined as 1.

Gene	Clone	Species	Stages	Anatomy
tbxt.S		laevis	NF stage 10	mesoderm

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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).

Gene	Clone	Species	Stages	Anatomy
tbxt		xenopus	NF stage 10.5	mesoderm , blastopore

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Figure 2. Specific Inhibition of Mesodermal Differentiation by XFDL(A) q-PCR analysis for Xbra (top), Mix.2 (middle), and Sox17α (bottom) expression. The four animal blastomeres at the 8-cell stage were injected with control mRNA (400 pg/cell; lanes 1 and 2) or 100 pg/cell (lane 3), 200 pg/cell (lane 4), or 400 pg/cell (lane 5) of XFDL mRNA. Animal caps (excised at stage 8.5) were then cultured without (lane 1) or with 5 ng/ml Activin (lanes 2–5) until siblings reached stage 11.(B) q-PCR analysis for Sox2 expression. Chordin (50 pg/cell; lanes 2–5) and increasing amounts of XFDL mRNAs were coinjected and analysis was performed as in (A).(C–R) In situ hybridization analysis of the embryos injected with XFDL mRNA. Control embryos (C, E, G, I, K, M, O, and Q) or embryos given XFDL mRNA (400 pg/cell) by injection at the 4-cell stage (D, F, H, J, L, N, P, and R) were harvested at stage 10.5 (C–L and O–R) or stage 11.5 (M and N) and analyzed by the indicated probes. Arrowheads, the dorsal and ventral blastopore lips; bracket, the marginal zone (see Xbra expression in panel C).(S and T) Embryos injected with SalF (400 pg/cell; S) or Tsh3 (400 pg/cell; T) were analyzed at stage 10.5 by in situ hybridization using the Xbra probe.