FIG 2.

HCMV pTRS1 or pIRS1 is necessary to antagonize PKR, maintain infected-cell protein synthesis, and inhibit stress granule formation. (A) Control cells (Scr) or shIRS1-HFs (shIRS1) were infected with either wild-type virus or HCMVΔTRS1 at an MOI of 3. Cells were harvested at 24 h after infection and analyzed by Western blotting (n = 3). Asterisks indicate nonspecific background bands. (B) Cells were infected as described for panel A, and the amount of radiolabeled amino acids incorporated into acid-insoluble protein in 30 min was quantified at 24 h after infection (n = 3; P < 0.05). Filled bars indicate wild-type infection, and open bars indicate HCMVΔTRS1 infection. (C) Control cells or shIRS1-HFs were infected with HCMVΔTRS1, and the formation of G3BP1 puncta was measured by indirect immunofluorescence at 24 h after infection. A representative image from one of three independent experiments is shown.