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Figure 2 | Retrovirology

Figure 2

From: The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu

Figure 2

Comparison of endogenous BST-2 in HeLa cells to BST-2 expressed in transiently transfected 293T cells. (A) 293T cells were transfected with wt BST-2 (lane 3). A mock transfected culture from HeLa (lane 1) and 293T cells (lane 2) was analyzed in parallel. Whole cell lysates were processed for immunoblotting as described in Methods. The arrow identifies a BST-2 species in transfected 293T cells not seen in HeLa cells. (B & C) Endoglycosidase analysis of transiently expressed BST-2. 293T cells were transfected with pcDNA-BST-2. BST-2 was enriched by adsorption to either datura stramonium lectin resin (DS lectin) (B) or Concanavalin A resin (ConA) (C) as described in Methods. DS lectin or ConA bound proteins were either left untreated (lanes 1 & 4) or treated with endoglycosidase H (EndoH) (lanes 2), Peptide: N-Glycosidase F (PNGase) (lanes 3), or endo-β-galactosidase (EndoB) (lanes 5) as described in Methods. Proteins were visualized by immunoblot analysis using a BST-2 specific antibody. (D) HeLa extracts were adsorbed to DS lectin (lane 2) and ConA resin (lane 3) as described for panels B & C. Total input lysate is shown in lane 1. A high mannose form of endogenous BST-2 was enriched on the ConA resin.

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Retrovirology

ISSN: 1742-4690

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