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. 2007 Mar 21;2(3):e316. doi: 10.1371/journal.pone.0000316

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Demeny et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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The nuclear localization of TAF8 lacking the NLS [TAF8(1-294] depends on its in vivo interaction with SPT7L. HeLa cells were co-transfected with the indicated CFP- and YFP-containing expression vectors and localization of the expressed proteins were visualised by fluorescence microscopy. The images shown in each panel are representative of all the transfected cells. (B) Sensitized emission of YFP fusion proteins due to FRET was measured in two different experiments in the nucleus of 25 individual HeLa cells transfected with the indicated combinations of vectors expressing YFP and CFP fusion proteins. The mean value of FRET efficiency (in%) over the entire nucleus in each cell was calculated as described in the Materials and Methods. A threshold was set to 5%, above the highest value of the negative control CFP/YFP (see horizontal line in each graph), and for the other pairs only values above this level were averaged. The average value of the negative control is 1.03%. The average value for each pair is the following: CFP-TAF8/YFP-TAF10 = 16.9%; CFP-TAF8/YFP-SPT7L = 15.8%; CFP-TAF6/YFP-TAF9 = 27.1%. Note that the scale for the TAF6/TAF9 is different.