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Figure 1


Fig. 1. Xenopus NEDD1 (XNEDD1) localizes to centrosomes and spindle microtubules. (Aa) Western blot of mitotic (M) and interphase (I) Xenopus egg extracts probed with antibodies raised against the C-terminal half of NEDD1 (amino acids 301-655). The position of molecular mass markers is indicated on the left. (b) Western blots of mitotic or interphase extracts treated with (+) alkaline phosphatase (AP) or control buffer (–). (B) Immunofluorescence micrographs of `cycled' microtubule asters (top panels) or spindles (bottom panels) induced by addition of sperm chromatin to egg extracts. Overlays show XNEDD1 in green and {alpha}-tubulin in red. Scale bar, 10 µm. (C) Immunofluorescence micrographs of sperm tips (top panels) incubated in egg extract in the presence of nocodazole to prevent microtubule assembly, or sperm-induced asters (bottom panels) triple-labeled with antibodies against XNEDD1 or {gamma}-tubulin, or stained for DNA, as indicated above the panels. Scale bars, 1 µm (top panels) and 10 µm (bottom panels). Overlays show XNEDD1 in green, {gamma}-tubulin in red and DNA in blue. (D,E) Xenopus tissue culture cells at various cell cycle stages (indicated on the left) triple labeled for XNEDD1, {alpha}-tubulin or {gamma}-tubulin and DNA, as indicated above the panels. Overlays in D show XNEDD1 in green, {alpha}-tubulin in red, DNA in blue. Overlays in E show {gamma}-tubulin in green, XNEDD1 in red, DNA in blue. Scale bars, 10 µm.