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. 2020 Apr 14;295(21):7327–7340. doi: 10.1074/jbc.RA120.013280

Figure 6.

Figure 6.

The cytoplasmic tail, GPI-anchor, and dimerization of human tetherin are required for ATP6V0C-mediated tetherin stabilization. A, schematic representation of tetherin, indicating that HA-tagged CT, TM, and CC domain contain glycosylation (NN) and dimerization (CCC) motifs and GPI-anchor (modified from Ref. 15). This image was originally published in Cell. Perez-Caballero, D., Zang, T., Ebrahimi, A., McNatt, M. W., Gregory, D. A., Johnson, M. C., and Bieniasz, P. D. Tetherin inhibits HIV-1 release by directly tethering virions to cells. Cell. 2009; 139:499–511. © Elsevier. B and C, 293T cells were transfected with vectors expressing HA-tagged tetherin with or without FLAG-tagged ATP6V0C expression vector. Tetherin variants tested include a cytoplasmic tail deletion mutant (delCT), a GPI-anchor deletion mutant (delGPI), a nonglycosylated mutant (NN), and a nondimerizing mutant (CCC) (B) and Agm tetherin (Agm), Rhesus tetherin (Rh), and a human tetherin containing the Agm (Hu-Agm) or Rh (Hu-Rh) transmembrane domains (C). One day post-transfection, cell lysates were prepared and immunoblotted with anti-HA to detect HA-tagged tetherin, anti-FLAG to detect FLAG-tagged ATP6V0C, or anti-tubulin antisera. Mobility of molecular mass standards is shown on the right of the blots.