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2O and reprecipitated with acetone several times to remove unincorporated radioactivity before loading onto SDS-polyacrylamide gels for analysis. Initial labeling experiments with 25 μCi of added cholesterol resulted in reduced incorporation by a factor of ∼10.
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2O, and extraction of the hydrophobic radioactivity with ether. After evaporation of the ether, the sample was resuspended in isopropanol and applied to the C18 column for analysis (20).
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4 dpm by excision and scintillation counting of an Hh-Np gel band.
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We are grateful to A. Woods and D. Raben for use of equipment and for helpful advice. We thank J. Nathans, D. Leahy, and T. Tanaka-Hall for comments on the manuscript. P.A.B. is an Investigator of the Howard Hughes Medical Institute.
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