Fig. 1. Claspin is phosphorylated at theMBT. (A) Temporal expression of ClaspinmRNA. RT-PCRwas performed on RNA isolated frommature oocytes and stagedembryos (Nieuwkoop and Faber, 1956) using primers specific for Claspin and actin as a control. (B) Spatial expression of Claspin protein. Whole-mount immunohistochemistry was performed with embryos at indicated stages using anti-Claspin serum(a) and preimmune serumas a control (f).An and Veg denote a viewfromanimal and vegetal poles, respectively.Unlabeled views are from the animal pole. (C) Temporal expression of Claspin protein. Western blotting with anti-Claspin, anti-phospho Chk1, anti-Cdk1 or anti-phospho Cdk1 (Y15) antibodies was performed on extracts from mature oocytes and staged embryos (first row), or in amore detailed time course, fromearly blastula (stage 7) to early gastrula (stage 10) embryos at indicated times after fertilization (second through fifth rows). Proteins equivalent to those fromhalf an embryo were analyzed. (D) Fertilized eggs (stage 1) and late blastula embryos (stage 9) were collected. Someembryoswere treatedwith hydroxyurea (HU) fromthe 2-cell stage andwere collected at the late blastula stage. The embryo extractswere subject to immunoprecipitationwith an anti-Claspin antiserum. Immunoprecipitates were treated without (−) or with (+) lambda phosphatase. Embryo extracts (left panel) and immunoprecipitates (right panel) were subjected toWestern blotting with an anti-Claspin antibody. -=unphosphorylated Claspin, and P=phosphorylated Claspin.
Image published in: Gotoh T et al. (2011)
Copyright © 2011. Image reproduced with permission of the Publisher, Elsevier B. V.
| Gene | Synonyms | Species | Stage(s) | Tissue |
|---|---|---|---|---|
| clspn.S | claspin, xClaspin | X. laevis | Sometime during NF stage 7 to NF stage 11 | animal cap vegetal pole |
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