Fig. 2. Depletion of xMIDs causes neural tube defects. (A) The complementary sequence of the xMID-Mo compared with xMID1 and xMID2. The ATG codons for the first methionine are underlined. (B) Western blot analysis of the N-terminal domain of the xMID proteins, tagged with Venus at the C-terminus (xMID1-Vns, xMID2-Vns, 250 pg), probed with an anti-GFP antibody. The xMID-Mo (17 ng) did not block translation of the mRNA for Venus (Vns, 100 pg) or the mRNA for xMID1-Venus with six silent mutations in the Mo recognition site (mut-xMID1-Vns, 250 pg). (C,D) Dorsal views of the unilaterally injected morphants; anterior is to the top. Control-Mo (C) or xMID-Mo (D) was injected into the right dorsal blastomere at the four-cell stage. Dashed lines indicate the boundaries between the neural and non-neural ectoderm. The black bracket marked by an asterisk indicates the distance between the neural folds in a rescue experiment. (E) Dorsal view of bilaterally injected xMID morphants at stage 20; anterior is to the top. (E′) Higher magnification view of the area marked by the bracket in E. (F,G) Transverse sections through the neural plate of Xenopus embryos with unilateral injection of control-Mo (F) or xMID-Mo (G). Dashed lines indicate the outlines of neural tissues, and brackets show the distance between the neural folds in a rescue experiment. (H) Average distance between the neural folds of unilaterally injected xMID morphants were dose-dependently reduced by the Mo-insensitive xMID1 (50, 100, 200, 500 pg), xMID2 (50, 100, 200, 500 pg), or xMID1+2 (25, 50, 100, 250 pg each) mRNAs. The number of embryos examined is indicated above each bar.
Image published in: Suzuki M et al. (2010)
Copyright © 2010. Image 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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