Figure 5.
ATP6V0C overexpression stabilizes tetherin. A, 293T cells were transfected with vectors expressing HA-tagged tetherin without or with FLAG-tagged ATP6V0C expression vector. 24 h post-transfection, cells were labeled with [35S]Met/Cys for 30 min and then chased for 0, 0.5, 1, 2, or 4 h in unlabeled medium. Cell lysates were prepared and immunoprecipitated with anti-HA antibodies and analyzed by SDS-PAGE followed by fluorography. Positions of 26-kDa (glycosylated) and 23-kDa (nonglycosylated) tetherin are indicated. B, the levels of radiolabeled 23- and 26-kDa tetherin species were quantified at each time point from three independent experiments ± S.D. (error bars). Tetherin level at time zero was taken as 100%. The p values (two-tailed paired t test) are calculated for each time point. *, p < 0.05; **, p < 0.02. The half-life (t½) of tetherin under each condition is indicated.