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Experiments using live cell 3‐color Förster (or fluorescence) resonance energy transfer (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...
IQGAP1 stimulates branched actin filament nucleation by activating N‐WASP, which then activates the Arp2/3 complex. N‐WASP can be activated by other factors, including GTP‐bound Cdc42 or Rac1, which also bind IQGAP1. Here we report the use of purified proteins for in vitro binding and actin polymerization assays, and Förster (or fluorescence) resonance energy transfer (FRET) microscopy of cultured...
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