Journal: Virology

Article Title: Tetherin restricts HSV-2 release and is counteracted by multiple viral glycoproteins.

doi: 10.1016/j.virol.2014.11.005

Figure Lengend Snippet: Fig. 4. Tetherin is highly likely to be incorporated into HSV-2 virions. (A) HSV-2 virion preparations were fractioned via a sucrose gradient cushion (10–50%) to separate cellular membrane fragments and defective viral particles from intact infectious virions. The representative viral and cellular proteins were assessed by western blot. One representative experiment out of three is shown. (B) Immunofluorescence analysis of HSV-2 virions. Virions harvested from HSV-2 infected HeLa cells were pelleted and applied to poly-L-lysine-coated coverslips prior to immunofluorescence analysis using anti-ICP5 and anti-tetherin antibodies. The FITC-positive dots were positive for tetherin and the Cy3-positive dots were positive for HSV-2 ICP5. Representative fields observed in two experiments are shown. Scale bars represent 500 nm.

Article Snippet: Cells were incubated for 1 h at 37 1C with a mouse monoclonal antibody against FLAG (F1804; Sigma) at a dilution of 1:200, gB (mouse monoclonal antibody; ab6506; Abcam) at a dilution of 1:200, gD (mouse monoclonal antibody; sc-58154; Santa Cruz) at a dilution of 1:50 or HSV-2 (sheep polyclonal antibody; PAB13979; Abnova) at a dilution of 1:200, followed by an incubation for 1 h at 37 1C with a FITC-conjugated goat anti-mouse secondary antibody (Boster, China) at a dilution of 1:100, a Cy3-conjugated goat antirabbit secondary antibody (Boster, China) at a dilution of 1:100 or a FITC-conjugated rabbit anti-goat secondary antibody (Boster, China) at a dilution of 1:100 in PBS-2% (w/v) BSA.

Techniques: Membrane, Western Blot, Infection