Fig 6.

Immediate-early expression of Us11 inhibits autophagy during infection with a recombinant virus lacking ICP34.5. (A) Immunoblot analysis of the Us11 protein in HeLa cells at 18 h p.i. (B) Immunoblot analysis of phosphorylated eIF2α protein in HeLa cells. Cells were infected with HSV-1, R3616, or R5104 at an MOI of 1 and transfected with poly(I · C) (5 μg/ml) for the last 4 h of infection. Total eIF2α blotting was used as a loading control. (C) Immunoblot analysis of LC3 protein in HeLa cells infected with HSV-1, R3616, or R5104 at an MOI of 1 at indicated times of infection. (D) Representative images of infected GFP-LC3 HeLa cells at 18 h p.i. Infected cells were visualized by an anti-ICP0 antibody, and GFP-LC3-positive cells with GFP-LC3 dots were quantified over the time course of infection. (E) Representative images of infected GFP-LC3 HeLa cells at 18 h p.i. when treated with poly(I · C) to induce autophagy. GFP-LC3-positive cells with GFP-LC3 dots were quantified in ICP0-positive cells. The results are the means of three independent experiments; 100 cells were analyzed per assay. **, P < 0.01 (t test). Scale bar, 10 μm.