Recapitulation of gro29 selection.

<p>Recapitulation of gro29 selection.</p

Co-localization of autophagosomes and viral capsids in HSV-1 infected gro29 cells.

<p>L/GFP-LC3 (A) and gro29/GFP-LC3 (B) cells were infected with HSV-1 mRFP-VP26 at an MOI of 10. At 6, 10 and 24 h post infection, the infected cells were fixed and the nuclei were stained with Hoechst. Arrowheads indicate areas in which the mRFP and the EGFP signals co-localize. Insets in the merged panels are magnified regions corresponding to the arrowheads. Confocal images are representative of three independent experiments.</p

Phosphorylation status of eIF2α in L and gro29 cells.

<p>L and gro29 cells, cells were mock-infected for 6 h, mock infected for 6 h then treated with 0.5 mM sodium arsenite for 30 min to stimulate eIF2α phosphorylation, or infected with HSV-1 at an MOI of 10 for 6 h. Equal volumes of whole cell lysate were electrophoresed through 10% polyacrylamide gels and transferred to PVDF membranes. Membranes were probed with antisera indicated on the left.</p

Induction of autophagy in L cells does not reduce the production of intracellular HSV-1.

<p>L (L) and gro29 (g) cells were pre-incubated for 1 h and maintained under nutrient rich (DMEM/10%FBS) (+) or nutrient deprivation (DMEM without FBS) (−) conditions and challenged with HSV-1 at an MOI of 1. At the indicated times post infection the cell associated virus was collected and titred on Vero cells. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042636#s2" target="_blank">Results</a> are from three independent experiments, with S.D. indicated.</p

Time course of gro29 cell survival and infection.

<p>(A) gro29 cells were infected with HSV-1 Us2-GFP at an MOI of 30 at 24 h post infection. Representative phase contrast (i) with corresponding GFP fluorescence (ii) images are shown. Phase contrast images of mock infected gro29 cells at 24 h (iii) and 192 h (iv). (B) The amount of virus produced from HSV-1 infected gro29 cells in the absence (grey line) or presence (black line) of a low pH wash every 24 h. At the indicated times post infection total infectious virus was quantified by titration on Vero cells and calculated as the number of plaque forming units (PFU) per mL. The sensitivity of the assay was 10¹ plaque forming units and is indicated by the thin horizontal grey line. (C) Time course of infected gro29 cells in the absence (No Wash) or presence (Plus Wash) of a low pH citrate buffer wash every 24 h. Representative phase contrast (PC) with corresponding GFP fluorescence images are shown.</p

gro29 cells accumulate virions in vesicles reminiscent of autophagosomes.

<p>At 18 h post infection, HSV-1 infected L (A) and gro29 (B) cells were harvested for electron microscopy. Representative electron micrographs are shown. Arrowheads indicate virions, while arrows identify cytoplasmic material within the vesicles. Inset in (A) shows a mature HSV-1 virion. Insets in (B) show a mature virion, an enveloped empty capsid and a non-enveloped nucleocapsid containing DNA (left to right).</p

Additional file 1: of Proteomic analysis of HIV-1 Gag interacting partners using proximity-dependent biotinylation

Proteins exclusively biotinylated by BirA*-Gag with known HIV-1 interactions. (PDF 96 kb