a 3x repeated nuclear localization signal and expressed in HeLa cells. For i-l bar is 10 µm and the fluorescence intensity was pseudo-colored between minimum and maximum fluorescence according to the mScarlet lookup table indicated in l. Micrographs shown in i-l are representative of at least 4 images made from 2 independent transfections.. Average FRET spectra from HeLa cells expressing
-RFP fusion proteins. All spectra were normalized to the unquenched donor and is calculated by Qs=E·Qa, in which Qa is the acceptor quantum yield of fluorescence. Qs is not dependent on the emission spectra and is proportional to the integrated acceptor sensitized emission signal. Each data point represents quantitative unmixing analysis of a single cell spectrum, 18–25 single cell spectra were recorded for each condition..