FIGURE 4.

Strategy of genetic screen. A, schematic of the genetic screen to identify host factors that rescue RTV-resistant HIV replication. B, Western blot analysis of WT HIV-RFP (R1) and HIV PRI54V/V82A-RFP (R2) reporter virus proteins compared with the protein profile of WT HIV. Defective cleavage of Gag in the HIV PRI54V/V82A-RFP reporter virus is indicated by the presence of p25 and p8 proteins. Panels represent individual viral proteins of the Pol polyprotein, accessory gene products, and surface (gp120) and transmembrane (gp41) Env proteins in mature virus particles. Shown are representative data from one of three independent experiments. C, Jurkat T cells infected with the HIV PRI54V/V82A-RFP reporter virus were sorted by flow cytometry and gated based on the intensity of red fluorescence. In the second round of FACS, total RFP cells were gated into two populations, and cells with the highest fluorescence intensity (0.01–0.1%) were sorted. SSC-A, side scatter area.