Ubulin-binding domain (TBD) are represented as grey and black boxes, respectively. Schematic representation of the Pr55/Pr55BRET assay. This assay is used as a sensor of Pr55multimerization. 293T cells were transfected with constant amounts of pCMV-Pr55-luc and increasing amounts of pCMV-Pr55-YFP. A constant amount of a third plasmid expressing Stau1-HAor Stau1-HAwas included in the transfection procedure. luc activity as well as transmitted and total YFP activities was measured. BRET ratios were plotted in function of their corresponding total YFP/luc ratio which allows us to compare BRET ratios at the same relative expression levels of Pr55fusion proteins. This figure is representative of four independent experiments. Cells corresponding to the four last points of each curve from Figure 4B were lysed. Cell lysates were analyzed by Western blotting using anti (α)-HA antibodies for their content in over-expressed Stau1 proteins. *: Non-specific labelling typically obtained with the anti-HA antibody. BRET ratios were compared at comparable total YFP/luc ratio. The BRET ratio corresponding to the pr55fusions expressed alone was arbitrarily set to 1. The BRET induction levels were then determined and are shown in the graph. These results are representative of 4 experiments.<p><b>Copyright information:</b></p><p>Taken from "The host protein Staufen1 interacts with the Pr55zinc fingers and regulates HIV-1 assembly via its N-terminus"</p><p>http://www.retrovirology.com/content/5/1/41</p><p>Retrovirology 2008;5():41-41.</p><p>Published online 22 May 2008</p><p>PMCID:PMC2409373.</p><p></p
