Figure 7
From: The Vpu-interacting Protein SGTA Regulates Expression of a Non-glycosylated Tetherin Species
Inhibition of ER-associated degradation has no effect on the accumulation of the non-glycosylated tetherin species.
(A) 293T cells were not transfected (none) or transfected with non-targeting (NT) or Bag6 shRNA constructs. One day later, the shRNA transfection was repeated. The cells were then transfected with vectors expressing HA-tagged tetherin, FLAG-tagged SGTA and Vpu. One day later, cells were lysed and subjected to western blotting with anti-HA, anti-Bag6, anti-Vpu and anti-FLAG antibodies. (B) 293T cells were transfected with the vector expressing HA-tagged tetherin with or without Vpu and FLAG-tagged SGTA expression vectors. Eight h post-transfection, cells were treated or not treated with kifunensine for 16–18 h and cells were lysed. Cell lysates were subjected to immunoblotting with anti-HA antibodies to detect HA-tagged tetherin, anti-FLAG antibodies to detect FLAG-tagged SGTA or anti-Vpu antisera. The location of the non-glycosylated, 23-kDa tetherin species is indicated by the arrow. (C) the intensity of non-glycosylated, 23-kDa tetherin species in lanes 5 and 6 was quantified from four independent experiments.
