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Effect of the H66N change on the shedding of gp120 in the cold. Radiolabeled HIV-1 bearing the ADA-N197S and ADA-H66N-N197S envelope glycoproteins (left) or KB9 and KB9-H66N envelope glycoproteins (right) was incubated on ice for 60 h or kept at −80°C. Viruses were pelleted and resuspended in radioimmunoprecipitation assay buffer. Viral pellets and supernatants were precipitated by a mixture of sera from HIV-1-infected individuals to determine the content of gp120 and p24 Gag proteins.
