TSG101 binds the class II FIPs.

<p>(<b><i>A</i></b>) Yeast two-hybrid analysis of the interaction between the indicated proteins. Protein-protein interactions were determined by the ability of the transformed yeast to grow on minimal medium lacking tryptophan, leucine and histidine (<i>His−</i>). <i>EV</i>, empty vector. (<b><i>B</i></b>) Co-immunoprecipitation analysis of the ability of Xpress-FIPs to co-immunoprecipitate GFP-TSG101 in HeLa cells using an anti-Xpress antibody (<i>SM</i>, starting material; <i>IP</i>, immunoprecipitate). Co-immunoprecipitated proteins were revealed using an anti-GFP antibody. GFP-empty vector (<i>EV</i>) was used as a control. SM load was 3.33%. (<b><i>C</i></b> and <b><i>D</i></b>) HeLa cells were transfected with constructs encoding the indicated proteins. At 16–18 hours post-transfection, cells were processed for immunofluorescence microscopy and immunostained with an anti-Xpress antibody. Cells expressing relatively low levels of the GFP-TSG101 protein are shown in <i>D</i>. DAPI was used to visualise the nuclei. Images were acquired by confocal microscopy. Insets illustrate the midbody region of dividing cells at 2.5× higher magnification. Scale bar indicates 10 µm. Data are typical of at least three independent experiments.