XB-IMG-156425
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Fig. 4.
Injection of embryos with either Xclaudin 1 MO or mRNA inhibits convergent extension movements. (A) Two dorsal blastomeres of 4-cell stage embryos, onto their equatorial region were injected with the Xclaudin 1 mRNA (500 pg; n = 28) or the Xclaudin 1 MO (20 ng; n = 30) separately, or together (mRNA, 250 pg, MO, 20 ng; n = 31). Dorsal marginal zone was dissected at early gastrula (st.10.5) and then cultured until stage 18. The elongation of explants was inhibited by injection with either Xclaudin 1 mRNA or Xclaudin 1 MO. The inhibition was partially rescued by co-injection of Xclaudin 1 mRNA. (B) We measured length/width ratio of injected DMZ explants in DMZ elongation assay. n, total number of explants. Error bars indicate means ± SD. (C) RT-PCR analysis showed that injection with either Xclaudin 1 mRNA (250 pg, 500 pg and 1 ng), or Xclaudin 1 MO (5, 10 and 20 ng) do not affect organizer cell fate specification from DMZ explants tissues. The expression of Chd, Gsc, Wnt11 and Xbra was analyzed using primers specific for them. EF1α is a loading control. (−) RT, minus reverse transcription; controls sample. (D) Whole-mount in situ hybridization analysis was performed after injection of either Xclaudin 1 mRNA (1 ng/embryo) or MO (40 ng/embryo) into equatorial regions of two blastomeres at 2-cell stage. Mesodermal marker Xbra and dorsal mesodermal genes Chd and Gsc were used in WISH. Neither Xclaudin 1 mRNA nor Xclaudin 1 MO changed the expression of the mesodermal fate markers. Image published in: Chang DJ et al. (2010) Copyright © 2010. This image is reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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