Xenbase Image ID: 134962

Fig. 1. Knockdown of ext1 results in gastrulation defects. (A) Embryos were injected with 20 ng of either the EXT1 MO or the mispair MO and lysed at st. 10.5 in preparation for immunoblotting to detect Ext1 and a-tubulin (loading control). Ext1 protein levels are decreased in EXT1-MO injected embryos. (B) Quantitative comparison of Ext1 protein accumulation. Relative Ext1 levels were normalized with a-tubulin. Bars represent mean ± S.D. N=3 independent experiments. (C) Midgastrula EXT1 MO-injected embryos exhibited incomplete blastopore formation and partial endoderm protrusion, whereas the mispair MO-injected embryos had a normal blastopore. (D,E) Embryos were injected with 20 ng of mispair or EXT1 MO at either the 2- cell stage or in either dorsal or ventral cells at the 4- cell stage; the phenotypes were compared at midgastrula (St. 10.5). A higher percentage of blastopore closure defects (D) or gastrulation arrest (E) were observed in EXT1 MO-injected embryos, irrespective of the stage or location of injection. N = 10 experiments, with a total of 400 embryos for each condition. (F) The phenotypes of embryos with head and tail defects in ext1 morphants. (G) The frequency of tail defects in ext1 morphants. Total number of tadpoles was 213. Bars present meant ± S.D. N=7 independent experiments. (H) The survival rates for EXT1 MO and EXT1 MIS embryos. Bars represent mean ± S.D. N=7 independent experiments (20-60 embryos/injection). (I) Blastopore closure defects and arrested gastrulation in ext1 morphants were partially rescued by coinjection with ext1 mRNA. Bars present meant ± S.D. N=5 independent experiments. (J) Embryos injected with 1ng of b-galactosidase or ext1 mRNA were collected and phenotypes were compared at gastrulation stage. The embryos overexpressing ext1 had higher frequencies of blastopore defects and gastrulation arrest compared to controls. Bars present meant ± S.D. N=5 independent experiments. * p < 0.05. Image published in: Shieh YE et al. (2014) Copyright © 2014. Reproduced with permission of the Publisher, University of the Basque Country Press. Larger Image Printer Friendly View

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