Journal:
Article Title: A Bipartite Membrane-Binding Signal in the Human Immunodeficiency Virus Type 1 Matrix Protein Is Required for the Proteolytic Processing of Gag Precursors in a Cell Type-Dependent Manner
doi:
Figure Lengend Snippet: Defect in proteolytic processing of HIV-1 Gag precursors in Myr− CD4+ T cells. Myr+ and Myr− CD4+ T-cell lines were generated as described in Materials and Methods. (A and B) Cell lysates from uninfected SupT-1 (lane 1), Myr−/SupT-1 (lane 2), and Myr+/SupT-1 (lane 3) cell lines were prepared by lysing of cells in radioimmunoprecipitation assay (RIPA) lysis buffer. The cell lysates were separated by SDS–12% PAGE and transferred to two nitrocellulose filters and then immunoblotted with an HIV-1-positive human serum (A) or with a monoclonal anti-RTp66/p51 antibody (B). (C) The supernatants from uninfected SupT-1 (lane 1), Myr−/SupT-1 (lane 2), and Myr+/SupT-1 (lane 3) cell lines were harvested after 48 h of incubation with fresh complete medium. The virion-associated proteins from harvested supernatants were concentrated by ultracentrifugation and analyzed by SDS-PAGE and immunoblotting with an HIV-1-positive human serum. (D) Cell lysates from uninfected H9 (lane 1), Myr−/H9 (lane 2), and Myr+/H9 (lane 3) cell lines were prepared by lysing the cells in RIPA lysis buffer. The cell lysates were separate by SDS–12% PAGE and the immunoblotted with an HIV-1-positive human serum.
Article Snippet: The CD4 + T-lymphoid cell line SupT-1 was also obtained from the American Type Culture Collection and was maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum and antibiotics.
Techniques: Generated, Radio Immunoprecipitation, Lysis, Incubation, SDS Page, Western Blot
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