Figure 3

IFITMs partly co-localize with HIV-1 Gag in virus-producing cells and coalesce with Gag into budding particles. A) To determine the intracellular distribution of IFITMs and Gag, HEK293T cells were transfected as mentioned above with the addition of a small amount of an HIV-1 Gag-GFP fusion protein (1/10 of WT Gag-Pol). Cells were then fixed 24 hours after and analyzed by confocal microscopy. Representative panels of more than 100 cells per condition are shown here. Scale bars: 10 μm. B) Co-localization corresponding to the yellow lines of (A) was performed using the Plot Profile tool in Image-J. C) The extent of reciprocal co-localization of IFITMs and Gag was quantified using the Manders overlap coefficient (Fuji image software). D) HEK293T cells co-transfected with IFITM3 and HIV-1 were analyzed by immuno-gold labeling with anti-p24 and anti-Flag antibodies (5 and 10 nm beads, respectively, as indicated). Negative controls consisted of cells transfected with control DNA. Scale bars: 200 nm. All panels present typical results obtained out of 2 to 3 independent experiments.