Xenbase Image ID: 158015

Figure 4. Ventx2 knockdown rescues the competence of MEK1-deficient cells to differentiate. (A) 4 cell embryos were injected in each cell with 50 pg Ventx2-Myc or 50 pg 2SAVentx2-Myc RNAs, and processed for WISH analysis at late gastrula stage 13 with pou5f3.2 probe. Stabilized 2SAVentx2 maintains ectopic pou5f3.2 expression. Top row dorsal view, bottom row lateral view. (B) Embryos injected as in (A), with or without 500 pg CA-Mk RNA per blastomere, were processed for WISH analysis at early gastrula stage 10.5 with t/bra probe. The number of embryos with ectopic t/bra expression is indicated in each condition. Stabilized 2SAVentx2 counteracts CA-Mk activity. Top row animal view, bottom row lateral view. (C) Sixteen-cell embryos were injected in one animal blastomere with 50 pg GFP-CAAX RNA, 25 ng Mk-MO and 7.5 ng Vx2-MO, as indicated. Next, these embryos were injected at blastula stage 8.5 with recombinant BMP4 (2 ng for xk81a1 induction, animal view), NODAL (5 ng for t/bra induction, lateral view; 50 ng for mixer (endoderm) induction, animal view), or NOGGIN (36 ng for sox2 induction, animal view) proteins into the blastocoel, collected at early gastrula stage 10.5, and processed for WISH. Mk-MO injected domains are indicated by white arrows. (D) Four-cell embryos were injected in each blastomere with 25 ng Mk-MO alone or with 7.5 ng Vx2-MO, collected at late gastrula stage 13 and processed for WISH with pou5f3.2 probe. (E) Embryos injected as in (D) were collected at tailbud stage 25, processed for WISH with sox2 probe, and scored. Ventx2 knockdown partially restores development of MEK1-deficient embryos. MEK1/Ventx2 double knockdown rescue assays were repeated five times. In A and C, the number of embryos exemplified by the photograph over the total number of embryos analyzed is indicated. Image published in: Scerbo P et al. (2017) © 2017, Scerbo et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Gene Synonyms Species Stage(s) Tissue pou5f3.2.L Oct-25, oct25, pou25, xoct-25, xoct25 X. laevis Throughout NF stage 13 neuroectoderm notoplate neural plate tbxt.L bra, brachyury, ntl, t, t-a, t-b, X-bra, Xbra, Xbrachyury X. laevis Throughout NF stage 10.5 mesoderm marginal zone krt12.4.L ck81, cytok, cytokeratin, E-keratin, epi-keratin, epidermal keratin, Epidermis-specific keratin, EpiK, epiker, EpK, epK, K81, xK81, xk81a, xk81a1 X. laevis Throughout NF stage 10.5 ectoderm sox2.L anop3, LOC108718087, mcops3, Sox-2, XLSOX-2, Xsox-2, XSox2 X. laevis Throughout NF stage 10.5 neuroectoderm Image source: Published Larger Image Printer Friendly View

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