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10.1002/cyto.a.22651 http://scihub22266oqcxt.onion/10.1002/cyto.a.22651 C4452401!4452401!25755111     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=25755111&cmd=llinks): Failed to open stream: HTTP request failed! 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Three-Color Confocal F�rster (or fluorescence) Resonance Energy Transfer Microscopy: Quantitative Analysis of Protein Interactions in the Nucleation of Actin Filaments in Live Cellsa |pdf=|usr=}}{{25755111}} gab.com Text Three-Color Confocal F�rster (or fluorescence) Resonance Energy Transfer Microscopy: Quantitative Analysis of Protein Interactions in the Nucleation of Actin Filaments in Live Cellsa JO: Cytometry A http://www.kidney.de/pget.php?&tw=1&pmid=25755111 #FOAvip Experiments using live cell 3-color FRET microscopy and corresponding in vitro biochemical reconstitution of the same proteins were conducted to evaluate actin filament nucleation A novel application of 3-color FRET data is demonstrated, extending the analysis beyond the customary energy transfer efficiency (E%) calculations.. MDCK cells were transfected for co-expression of Teal-N-WASP/Venus-IQGAP1/mRFP1-Rac1, Teal-N-WASP/Venus-IQGAP1/mRFP1-Cdc42, mCFP-Rac1/Venus-IQGAP1/mCherry-actin or CFP-Cdc42/Venus-IQGAP1/mCherry-actin, and with single-label equivalents for spectral bleedthrough correction. Using confirmed E% as an entry point, fluorescence levels and related ratios were correlated at discrete accumulating levels at cell peripheries. Rising ratios of CFP-Rac1:Venus-IQGAP1 were correlated with lower overall actin fluorescence, whereas the CFP-Cdc42:Venus-IQGAP1 ratio correlated with increased actin fluorescence at low ratios and was neutral at higher ratios. The new FRET analyses also indicated that rising levels of mRFP1-Cdc42 or mRFP1-Rac1 respectively promoted or suppressed association of Teal-N-WASP with Venus-IQGAP1. These 3-color FRET assays further support our in vitro results about the role of IQGAP1, Rac1 and Cdc42 in actin nucleation, and the differential impact of Rac1 and Cdc42 on the association of N-WASP with IQGAP1. Moreover, this work emphasizes the power of 3-color FRET as a systems biology strategy for simultaneous evaluation of multiple interacting proteins in individual live cells. Twit Text FOAVip Three-Color Confocal F�rster (or fluorescence) Resonance Energy Transfer Microscopy: Quantitative Analysis of Protein Interactions in the Nucleation of Actin Filaments in Live Cellsa ????Cytometry A http://www.kidney.de/pget.php?&tw=1&pmid=25755111 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25755111 #FOAvip English Wikipedia <ref>{{Cite pmid|25755111}}</ref> Three-Color Confocal F�rster (or fluorescence) Resonance Energy Transfer Microscopy: Quantitative Analysis of Protein Interactions in the Nucleation of Actin Filaments in Live Cellsa #MMPMID25755111 Wallrabe H; Sun Y; Fang X; Periasamy A; Bloom GS Cytometry A 2015[Jun]; 87 (6): 580-8 PMID25755111show ga Experiments using live cell 3-color FRET microscopy and corresponding in vitro biochemical reconstitution of the same proteins were conducted to evaluate actin filament nucleation A novel application of 3-color FRET data is demonstrated, extending the analysis beyond the customary energy transfer efficiency (E%) calculations.. MDCK cells were transfected for co-expression of Teal-N-WASP/Venus-IQGAP1/mRFP1-Rac1, Teal-N-WASP/Venus-IQGAP1/mRFP1-Cdc42, mCFP-Rac1/Venus-IQGAP1/mCherry-actin or CFP-Cdc42/Venus-IQGAP1/mCherry-actin, and with single-label equivalents for spectral bleedthrough correction. Using confirmed E% as an entry point, fluorescence levels and related ratios were correlated at discrete accumulating levels at cell peripheries. Rising ratios of CFP-Rac1:Venus-IQGAP1 were correlated with lower overall actin fluorescence, whereas the CFP-Cdc42:Venus-IQGAP1 ratio correlated with increased actin fluorescence at low ratios and was neutral at higher ratios. The new FRET analyses also indicated that rising levels of mRFP1-Cdc42 or mRFP1-Rac1 respectively promoted or suppressed association of Teal-N-WASP with Venus-IQGAP1. These 3-color FRET assays further support our in vitro results about the role of IQGAP1, Rac1 and Cdc42 in actin nucleation, and the differential impact of Rac1 and Cdc42 on the association of N-WASP with IQGAP1. Moreover, this work emphasizes the power of 3-color FRET as a systems biology strategy for simultaneous evaluation of multiple interacting proteins in individual live cells. �Three-Color Confocal F�rster (or fluorescence) Resonance Energy Transf(epmc).pdf DeepDyve Pubget Overpricing