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

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Figure 2. — ATP6V0C overexpression has no effect on HIV-1 release. A, 293T cells were transfected with WT or Vpu-defective (delVpu) pNL4-3 HIV-1 molecular clones with or without vectors expressing FLAG-tagged ATP6V0C and HA-tagged human tetherin. One day post-transfection, cell and viral lysates were prepared and subjected to Western blot analysis with HIV-Ig to detect the Gag precursor Pr55Gag (Pr55) and the CA protein p24 or with anti-FLAG to detect FLAG-tagged ATP6V0C, anti-HA to detect HA-tagged tetherin, or antisera against Vpu or tubulin. Mobility of molecular mass standards is shown on the right of the blots. B, virus release efficiency (VRE) was calculated as the amount of virion-associated p24 (CA) relative to total Gag in cell and virus. VRE was set to 100% for WT pNL4-3 in the absence of ATP6V0C and tetherin. Data shown are mean ± S.D. (error bars) from three independent experiments.