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. 2006 Dec 25;203(13):2887–2893. doi: 10.1084/jem.20061519

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — A3G restricts HIV-1 infection in iDC 293T. A3G-HA cells (A) or iDCs (B) were transfected with siRNA against A3G (Si-A3G) or siRNA control (Si-Ctrl). After two rounds of transfection, cells were analyzed by Western blot (A and B, left). The percentage of protein expression on Western blot was quantified by density of specific bands and normalized with Si-Ctrl (A and B, right). 293T-A3G-HA cells (C) or iDCs (D) transfected by Si-A3G and Si-Ctrl were infected with HIV-VSVG. GFP expression was followed by FACS (numbers are the percentage of GFP⁺ cells; C and D, left), and the fold enhancement of GFP expression was calculated by normalization with nontreated (NT) cells (C and D, right). A pool of five independent experiments is shown for A3G (±SEM), and two independent experiments are shown for A3F (±SEM). NI, noninfected. (E) A3G/3F induce G-to-A hypermutation of HIV genomes during iDC infection. Sequences represent individual HIV-1 reverse transcripts obtained after an 8-h infection of iDCs and are representative of the 7 hypermutated sequences (17%) that were obtained among 40 reverse transcripts. The standard HIV NL4.3 numbering is used.