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500 nM Gαi1-CFP and 900 nM YFP-RGS4 were excited at 433 nm and emission spectra measured with a cuvette-based PerkinElmer LS-55 spectrometer. In the presence of AMF, RGS4 and Gαi1 bind with high affinity, leading to FRET between their fluorescent tags and an increased FRET ratio ((525 nm emission)/(474 nm emission)) compared to GDP buffer.
