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Immunofluorescence staining of Xenopus embryos was done as described (8), with affinity-purified polyclonal antibody to cyclin E (9) and monoclonal antibodies to α tubulin or γ tubulin (Sigma). Confocal microscopy was performed on an MRC-600 system (Bio-Rad, Hercules, CA). The confocal images presented represent projections of Z-series scans.
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We thank F. J. Miller for help developing the observation chambers, C. Wilkerson and M. Cham for assistance in building the image acquisition computers, and R. Pope, T. Pederson, G. Witman, and B. Luna for comments on the manuscript Supported by the NIH (J.L.M. and G.S.), the Trustees of the Worcester Foundation (G.S.), and the Cabot Family Charitable Trust (G.S.). J.L.M. is an investigator of the Howard Hughes Medical Institute. E.H.H. is supported by an NIH postdoctoral training fellowship. The authors would like to dedicate this work to the memory of Dan Mazia.
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