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. 2009 Mar 13;5(3):e1000339. doi: 10.1371/journal.ppat.1000339

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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(A) First two upper left panels show a cell expressing HA-Bro_i (white). The next panel shows a reconstructed 3D view of a stack of Z sections from a cell expressing HA-Bro_i and Gag-GFP. Lower panels show a single Z section of the same cell showing colocalization of Bro_i and Gag-GFP at the plasma membrane. Bro_i (white) was stained with a mouse monoclonal anti-HA antibody and an Alexa 633-conjugated anti-mouse antibody. Nuclei were counterstained with DAPI (blue). F-actin was stained with Alexa 568-conjugated phalloidin (red) to delineate cells. The colocalization channel (yellow) of Bro_i and Gag-GFP was built using Imaris software. Scale bar = 5 µm. (B, C) Electron micrographs of 293T cells co-transfected with pNL4-3 wt and HA-Bro_i. (B) Arrested budding structures are indicated with black arrows. Two regions of interest in (a) and (b) show budding structures carrying electron-dense crescent-shaped material at a higher magnification. (C) HIV-1 budding structures tethered to the plasma membrane.