BST2/Tetherin enhances HCMV infection and release.

<p>A) A schematic representation of the BST2 constructs used in this study. B) Immunoblot for HCMV-IE1 or GAPDH of HFFs exposed to supernatants of HCMV-infected THFs expressing BST2-HA146 or vector control. Supernatants were obtained after infection with HCMV strains AD169 or Toledo (MOI = 3) for 72 hrs. C) Viral titers in supernatants of AD169-infected HFFs stably expressing the indicated BST2 constructs. Virus was titrated by end point serial dilution assay. D) Percentage of GFP positive cells of THF-HA146, THF-PCDH, HFF-HA146 and HFF-PCDH cells transfected with the indicated siRNAs 72 hours prior to infection with AD169-GFP for 24 h. GFP levels in GAPDH controls were set to 100%. E) Immunoblot for IE1 and GAPDH in HFFs, THFs and THF-HA146 cells transfected with the indicated siRNA 72 h prior to infection with HCMV Toledo for 8 h. F) Immunoblot for IE1, BST2 and GAPDH in HFF-HA146 cells transfected with indicated siRNAs prior to infection with HCMV AD169 at the indicated MOI for 8 h. For control 40 nM silencer negative control (Ambion) was used. The numbers below the blots in E) and F) show the relative IE1 band intensity compared to GAPDH (using ImageJ) normalized to control siRNA.