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

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Figure 4. — Tetherin co-immunoprecipitates with ATP6V0C independent of Vpu. 293T cells were transfected with vectors expressing FLAG-tagged ATP6V0C without or with HA-tagged tetherin and Vpu expression vectors (A) or HA-tagged tetherin without or with FLAG-tagged ATP6V0C and Vpu expression vectors (B). Twenty-four h post-transfection, cell lysates were prepared and immunoprecipitated with agarose beads conjugated with anti-FLAG (A) or anti-HA (B) antibodies. Complexes were washed, and both cell lysates and immunoprecipitated samples were subjected to immunoblotting with anti-FLAG, anti-HA, and anti-Vpu antibodies to detect FLAG-tagged ATP6V0C, HA-tagged tetherin, and Vpu, respectively. Mobility of molecular mass standards is shown on the right of the blots. The location of different species of tetherin and Ig light chain (IgL) is indicated by the arrows.