Fig. 1

Tetherin restricts cell-to-cell spread of HSV-2. HeLa cells were infected with HSV-2 using 0.0001PFU/cell. Two hours later, the virus inoculum was removed and cells were incubated in the medium containing anti-HSV-2 antibody a or incubated in the normal medium without anti-HSV-2 antibody b. After 2 days, the cells in a and b were fixed and stained for HSV-2 antigens. The peroxidase-conjugated secondary antibody and substrate were used to reveal HSV-2-infected cells. c The expression of total tetherin in HeLa cells pretreated with tetherin siRNA or control siRNA was analyzed by western blot where actin was used as a loading control. Molecular weight standards in kilodalton are shown on the left. d and e The morphology of HSV-2 plaques on HeLa monolayers pretreated with tetherin siRNA d or control siRNA e. Representative fields observed in four experiments are shown. f The morphology of HSV-2 plaques on HaCaT and ARPE-19 monolayers transfected with pcDNA3.1 or pBST2 using the infectious center assay as described in the Materials and Methods. Representative morphology of HSV-2 plaques on HaCaT and ARPE-19 monolayers are shown. Scale bars in all panels represent 100 μm. g Representative fields containing more than 10 plaques were chosen and the plaque areas were calculated. The plaque area of the pcDNA3.1-transfected cells was arbitrarily set at a value of 100 for comparison with that of the pBST2-transfected cells. Data shown are mean ± SD