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Fig. 2. Depletion of Ascl1 increases mesendoderm gene expression. (A-C) Western blot analysis of the efficacies of Ascl1 MOa (A), MOa2 (B) and MOb (C) in blocking the translation of synthetic reporter ascl1 mRNA with the wild-type 5′ UTR or with a mutated 5′ UTR. Doses of injected MOs: 50 or 100 ng. (D) Representative control and Ascl1 morphant embryos at different development stages. Experiment for each and every MO was repeated at least in three independent experiments, with embryos from one representative experiments shown in the figure. Numbers in the lower panels indicate incidence of morphological appearance resembling the embryos shown. (E,F) Control MO (cMO), Ascl1 MOa or MOb (E), or MOa2 (F) injected at one-cell stage and resultant morphants collected at stage 10.5 followed by qPCR examination of gene expression normalized to the levels of odc. **P<0.05, Student's t-test. (G) GO analysis of 384 upregulated genes in Ascl1 morphants against cMOs at stage 10.5-11. (H) Heat map presentation of the relative expression levels of a subset of VegT targets upregulated by Ascl1 depletion. (I) cMO or Ascl1 MOs (MOa+b) injected at one-cell stage and marginal zone explanted at the stage 8 and cultured to the equivalent stage 10.5 followed by qPCR examination of gene expression normalized to the expression levels of odc. Changes in expression of all examined genes relative to control are significant: Student's t-test, P<0.05. Image published in: Gao L et al. (2016) © 2016. 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 Larger Image Printer Friendly View |