Figure 1: HIV-1 blocks SG assembly irrespective of eIF2α-P.

HeLa (a) and Jurkat cells (c) were transfected with HIV-1 pNL4-3 and were exposed to PatA (middle panel) for 1 h or sodium Ars (bottom panel) for 45 min. Cells were stained for G3BP1, TIAR and Gag. Yellow arrows: mock-transfected cells. Red arrows: HIV-1-expressing cells. Scale bar, 10 μm. Magnifications of cells (c) on right show presence of SG indicated with green arrows. (b,d) Top: quantification of SG in mock-transfected (grey bars) or HIV-1 expressing (black bars) cells from a,c. Data are presented as mean±s.d. from three separate experiments with at least 100 cells each analysed. Bottom: cell lysates were analysed for eIF2α-p (Ser51), eIF2α, Gag and GAPDH. (e) HeLa cells expressing HIV-1 and G3BP1-GFP or TIA-1-GFP (0.5–2 μg) were processed for FISH/IF analyses and stained for Gag and vRNA. Scale bar, 10 μm. (f) Immunoblot analysis of whole-cell lysate from e is shown. (g,h) PBMCs from two HIV-1-infected subjects with different disease outcomes were subjected to stress and analysed by FISH/IF. Cells from long-term nonprogressor (LTNP; g) and typical progressor (h) were fixed and stained for G3BP1 and vRNA. Scale bar, 5 μm. Magnifications of cells on right show presence of SG indicated with green arrows. Right Panel: quantification of SG in uninfected (grey bars) or HIV-1-infected (black bars) cells from g,h. Data are presented as mean±s.d. from three separate experiments with at least 50 cells each analysed.