Fig. 3.

CT mutations increase surface expression of Envs in recombinant VACV-infected cells. HEK 293T cells were infected (MOI = 3) with the indicated Env-expressing recombinant VACV or parental VACV (empty). After 16 h, cells were stained for Env using (A) 2G12, an anti-gp120 mAb, or (B) 2F5, an anti-gp41 mAb under permeabilizing (total) or non-permeabilizing (surface) conditions (N = 3). In each experiment, VACV A33 protein staining was performed under permeabilizing conditions to control for transduction efficiency and to gate on infected cells. The MFI of Env and A33 stains are shown relative to parental Envs, with comparison groups separated by dotted lines. Asterisks indicate a significant fold difference (p < 0.05) in surface expression of WT and TM1 Envs by one-sample t-test on log-transformed data with Bonferroni correction.