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. 2009 Oct 28;106(49):20889–20894. doi: 10.1073/pnas.0907075106

Fig. 5.

Fig. 5.

Tetherin and SIVtan Env co-localize specifically in tubulo-vesicular structures. (A and B) 293T cells expressing HIV-1 Gag and X-THN were stained with anti-tetherin antibodies and 10 nm PAG. Tetherin is seen at the cell surface and on intracellular membranes which can be arranged in tight clusters of >0.5-μm diameter. (C and D) In 293T cells co-expressing HIV-1 Gag, X-THN, and HA-SIVtan Env, tetherin is still found on membranes in the Golgi area and on tight clusters of tubulo-vesicular membranes. (E–G) Staining with mouse anti-HA and 10 nm PAG shows that the HA-SIVtan Env protein is also on Golgi stacks, membranes in the Golgi area, and on tight clusters of tubulo-vesicular membranes, which can be expanded into large clusters (E). (H–J) Double labeling shows co-localization of HA-SIVtan Env and tetherin in tubulo-vesicular membranes. (I) Shows an enlargement of the area marked in H. Sometimes tetherin (PAG 10 nm, e.g., at the black arrows) can be seen in vesicles containing the HA-SIVtan Env (PAG 5 nm). (J) Tetherin (PAG 5 nm, e.g., at the black arrows) co-stains with HA-SIVtan Env (PAG 10 nm). No gold labeling was seen on untransfected cells. G, Golgi apparatus; m, mitochondrion; PM, plasma membrane. (Scale bars, 200 nm.) All images are representative of at least eight cells.