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. 2014 Dec 5;9(12):e114753. doi: 10.1371/journal.pone.0114753

Figure 6. Role of the membrane-proximal YXXΦ motif.

Figure 6

(A) Schematic representation of additional mutations within the cytoplasmic domain of gp41. Black boxes indicate location and type of potential trafficking motifs. Black arrows represent overlapping reading frames for rev, tat, and nef, respectively. (B and C) Incorporation of mutant Env into virions. Viruses were produced by transfection of HEK293T cells and virions were pelleted from the clarified supernatants. (B) gp160/gp120 (top panel) and p27 (bottom panel) were detected by Western blot using antibody 3.11H and 2F12, respectively. (C) Relative Env incorporation into virions. The ratios of gp120 to p27 were calculated by phosphorimaging analysis. Data are presented as the gp120:p27 ratio of mutant virions (Rn) relative to the gp120:p27 ratio of SIVmac239 wild-type (WT) virions (R239). (D) Infectivity of mutant viruses. Stocks were normalized for p27 content and used to infect C8166-45 SIV-SEAP cells with serial dilutions. SEAP activity 3 days after infection. Error bars indicate standard deviation of triplicates.