Figure 3. Mesodermal Conversion Induced by XFDL Knockdown in the Animal Cap(AâJ) 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 2â4) 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.
Image published in: Sasai N et al. (2008)
Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| tbxt.S | bra, brachyury, ntl, t, t-a, t-b, X-bra, Xbra, Xbrachyury | X. laevis | Throughout NF stage 10 | mesoderm |
| sox2.L | anop3, LOC108718087, mcops3, Sox-2, XLSOX-2, Xsox-2, XSox2 | X. laevis | Throughout NF stage 10 | ectoderm dorsal |
| sox17a.L | sox17 alpha, sox17-alpha, Sox17alpha, tSox17alpha, xsox17, xsox17a, Xsox17-alpha, Xsox17alpha, xSox17alpha1, xSox17alpha2 | X. laevis | Throughout NF stage 10 | endoderm upper blastopore lip |
| vegt.L | antipodean, brat, tVegT, vegt-a, vegt-b, xombi | X. laevis | Throughout NF stage 10.5 | mesoderm lateral ventral |
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