Skip to main content
. Author manuscript; available in PMC: 2016 Apr 25.
Published in final edited form as: J Mol Biol. 2015 Feb 7;427(6 Pt B):1413–1427. doi: 10.1016/j.jmb.2015.01.018

Figure 1.

Figure 1

Virus release and membrane binding of 29KE/31KE and compensatory mutants. (a) HeLa cells were transfected with the HIV-1 mutants indicated, labelled with 35S Met/Cys for 3 h, then cell and virus lysates were immunoprecipitated and separated by SDS-PAGE. Samples from three independent experiments were analyzed by fluorography. Virion capsid, expressed as a percentage of total Gag, was plotted, +/− S.E.M.. (b) HeLa cells were transfected with the HIV-1 mutants indicated (in a PR– virus), then labelled with 35S Met/Cys for 20 min. Cells were harvested by scraping, lysed by sonication and lysates placed at the bottom of a sucrose gradient. Following centrifugation, membrane-associated samples were collected from the top of the gradient, and soluble samples from the bottom. Gag was immunoprecipitated from these fractions and resolved by SDS-PAGE. Samples from four independent experiments were analyzed by fluorography. Gag percentage in the membrane-associated fraction was plotted +/− S.E.M. Flotation fractions were also analyzed by western blotting for transferrin receptor and GAPDH.