Figure 3
From: The Vpu-interacting Protein SGTA Regulates Expression of a Non-glycosylated Tetherin Species
Overexpression of SGTA in the presence of Vpu increases the levels of a non-glycosylated 23-kDa form of tetherin.
(A) 293T cells were transfected with WT or Vpu-defective (delVpu) pNL4-3 HIV-1 molecular clones and vectors expressing HA-tagged human tetherin with or without FLAG-tagged SGTA expression vector. One day posttransfection, cell and viral lysates were prepared and subjected to western blot analysis with HIV-Ig to detect the Gag precursor Pr55Gag (p55) and the CA protein p24, anti-FLAG to detect FLAG-tagged SGTA or anti-HA to detect HA-tagged tetherin or anti-Vpu antisera. Mobility of molecular mass standards is indicated to the right of the anti-HA blot. The location of the 23-kDa tetherin species is indicated by the arrow. (B) 293T cells were transfected with vectors expressing HA-tagged WT or glycosylation-defective tetherin mutants (N65A, N92A and NN-AA) with or without FLAG-tagged SGTA and Vpu expression vectors. One day posttransfection, cells were lysed and subjected to western blot analysis with anti-HA to detect HA-tagged tetherin or anti-FLAG to detect FLAG-tagged SGTA or anti-Vpu antisera. Molecular mass markers are shown on the right of the anti-HA blot. The location of the non-glycosylated, 23-kDa tetherin species is indicated by the arrow.
