Plasma membrane–connected CD36⁺ compartments are present in infected and uninfected macrophages. (A) Confocal micrographs of uninfected macrophages stained for the indicated antibodies. (B) Confocal sections of primary macrophages co-infected with HIV-1 Gag-iCherry ΔEnv and CD36-GFP lentiviral vector for 7 d, fixed, and stained for the indicated markers. In B and E anti-GFP antibodies were used to enhance the GFP signal. (C) Macrophages grown on fluorodishes with coordinates were infected with a lentiviral vector encoding CD36-GFP, together with SIV-VLP and, 5 d later, with HIV-1 Gag-iCherry ΔEnv. On day 7 p.i., cells were exposed to a 10 kD dextran–Alexa Fluor 647 and then immediately imaged by spinning disk microscopy to follow the indicated markers. The same fluorodishes were then embedded in epon resin and processed for EM. An overview of the macrophage imaged by EM is presented on the right. N = nucleus. (D) Magnification of the region boxed on the right, with viral budding profiles visible at the limiting membrane and both mature and immature viral particles. (E) Macrophages infected with the lentiviral vector encoding CD36-GFP for 7 d were fixed and stained for the VCC marker CD81. (F) Macrophages infected with the lentiviral vector encoding CD36-GFP for 12 d were exposed as in C to dextran–Alexa Fluor 647 and then immediately imaged by spinning disk microscopy. In A, B, and E merge images include DAPI staining in blue. Data shown are representative of at least two independent experiments. Bars: (A–C, E, and F) 10 µM; (D) 2 µM.