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. 2005 Apr 11;169(1):49–60. doi: 10.1083/jcb.200411118

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Copyright © 2005, The Rockefeller University Press

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Figure 3. — Characterization of Xenopus ZW10 and Rod. (A and B) Schematic alignment of Xenopus and human ZW10 (A) or Xenopus Rod_COOH (XL107l09) and human Rod (B). Amino acid positions as well as percentage identity and additional (*) similarity on the protein level are indicated. (C) Coomassie staining of purified recombinant X-ZW10 (X-Z) and X-Rod_COOH (X-R_COOH). (His)₆-tagged proteins were purified from insect cells and analyzed by Coomassie blue staining. (D) Immunoblot analysis of pAbs to X-ZW10 and X-Rod_COOH. 20 ng of recombinant protein (rec. prot.) and 1 μl CSF extract were analyzed by immunoblot with affinity purified anti–X-ZW10 (1348) or anti–X-Rod_COOH (1351). Position of the endogenous frog proteins in the CSF extract is indicated. Cross-reacting proteins are marked by asterisks. (E) Immunolocalization of X-ZW10 and X-Rod. Sperm nuclei replicated in cycled CSF extract were immunostained for X-BubR1 and X-ZW10 or X-Rod. DNA (DAPI) is in blue. Enlarged boxes show overlap of X-BubR1 and X-ZW10–X-Rod signals on a sister kinetochore pair.