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Fig. 5 (Right). Mesodermal genes were not influenced by inhibition of xGLUT1. (A) xGLUT1MO inhibited elongation of animal caps. 20 ng of xGLUT1MO or control MO was injected into the animal pole of an 8-cell stage embryo. At late blastula, animal caps were dissected and treated with 10 ng/ml of activin and cultured for 10 hours. Without activin treatment, caps remained round (A.a-c) and activin-treated caps dissected from control embryos (d) or standard control MO-injected embryos (e) became elongated in response to activin. However, the elongation of xGLUT1MO-injected caps was inhibited (A.f). (B) Expression of mesodermal marker genes in animal caps injected with xGLUT1MO. Animal caps were treated with activin for 3 hours and sampled for RT-PCR. Xbra, mix2 and Chd transcript levels were not influenced by xGLUT1MO injection. ODC was used as a loading control. (C) Whole-mount in situ hybridization (WISH) with late gastrula. 10 ng of xGLUT1MO and 100 pg of β-gal mRNA was coinjected into one side of the dorsal marginal zone of 4-cell stage embryos. Before WISH, the embryos were stained with Red-gal for cell lineage tracing. xGLUT1MO injection caused no change in xbra (a), chd (b), or gsc (c) expression. |
