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ATP and FRET—a cautionary note

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Figure 1: ATP concentration affects signal efficacy of CFP-YFP–based FRET sensors.
Figure 2: FRET sensors show ATP concentration–dependent YFP/CFP peak intensity ratio changes in permeabilized cells in vivo.

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Acknowledgements

This work was supported by ZonMw HORIZON Breakthrough Grant 050-71-019 and NKB-KWF grant 2002-2763; we thank L. He, St. Jude Children's Research Hospital, for kindly providing us with the CFP-TRAF2TD-YFP construct, and R. Lynch, University of Arizona, for helpful suggestions.

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Supplementary information

Supplementary Figure 1

Characteristics of the different sensors initially designed to monitor ATP concentrations. (PDF 219 kb)

Supplementary Figure 2

Effect of ATP on FRET ratio is not the result of a change in fluorescent properties of the CFP or YFP fluorophores. (PDF 2029 kb)

Supplementary Figure 3

The effect of [ATP] on the FRET ratio of the Xa protease cleavage site based control sensor is dependent on the MgCl2 concentration in the buffer. (PDF 723 kb)

Supplementary Video 1

Human skin fibroblasts showing the changes in the spectra of the XProt-sensor in time upon washout and addition of ATP. (AVI 9089 kb)

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Willemse, M., Janssen, E., Lange, F. et al. ATP and FRET—a cautionary note. Nat Biotechnol 25, 170–172 (2007). https://doi.org/10.1038/nbt0207-170

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