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. 2012 Mar 1;287(18):14837–14850. doi: 10.1074/jbc.M112.349928

FIGURE 1.

FIGURE 1.

Vpu-dependent ubiquitination of endogenously expressed BST-2. A, HeLa cells transfected with pHA::Ub and the indicated pNL4–3 proviral clones were treated ± the lysosomal inhibitor CMA. Lysates were made from these cells, BST-2 was immunoprecipitated from the lysates using an anti-BST-2 mAb (Chugai), and the final IP pellets were treated with peptide N-glycosidase F. Shown here is an immunoblot of those IPs probed with anti-BST-2 polyclonal antisera. B, the immunoblot shown in A was stripped and reprobed with an anti-HA mAb to detect the presence of HA-tagged ubiquitin conjugated to the immunoprecipitated BST-2 proteins. C, the immunoblot shown in A and B was stripped and reprobed with the anti-Ub mAb P4D1. D, an immunoblot of the input lysates used in A and B was probed with an anti-Vpu polyclonal antibody to show the level of Vpu expressed in these cells. E, the immunoblot (IB) shown in D was stripped and reprobed with an anti-actin antibody.