XB-IMG-45909
Xenbase Image ID: 45909
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Fig. 1. Smicl is a Smad-interacting protein that is expressed maternally and then zygotically during Xenopus development. (A) Amino acid sequence of XtSmicl, a novel Smad interacting protein. Conserved cysteines and histidines in the zinc fingers are underlined. (B) Smicl interacts with Smad3 and Smad4. HA-tagged XtSmicl was expressed in HEK293T cells with plasmids encoding Myc-tagged Smad1, Smad2, Smad3, Smad4 or Smad5 in the presence or absence of the appropriate constitutively active type I receptor. Samples were immunoprecipitated (IP) using an anti-Myc antibody and the presence of HA-XtSmicl was analysed by western blotting (W). (C) Smicl interacts with endogenous Smad3. HA-tagged XtSmicl was expressed in HEK293T cells in the presence or absence of caALK4. Samples were immunoprecipitated with anti-HA coupled beads and the presence of endogenous Smad3 was analysed by western blotting. (D) Expression pattern of Xenopus tropicalis Smicl studied by whole-mount in situ hybridisation. Stages (st) are indicated. a, animal; v, vegetal; D, dorsal; V, ventral; A, anterior; P, posterior; mes, mesoderm; ect, ectoderm. Smicl is expressed maternally and transcripts are particularly abundant in the animal hemisphere of the fertilised egg. These results were confirmed by in situ hybridisation experiments carried out on bisected embryos to facilitate penetration of the probe in the vegetal hemisphere of the embryo. In situ hybridisation using a sense probe revealed no background staining (data not shown). (E) Quantitative RT-PCR confirms the presence of high levels of Smicl RNA in the fertilised egg of Xenopus tropicalis. Significant zygotic expression of Smicl begins at tadpole stage 32. Image published in: Collart C et al. (2005) Copyright © 2005. 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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