Supplementary Figure 8: Visualizations of different averages for efficiencies according to different fluorophore dynamics.

(a) Dynamic average, which applies in the case of the fluorophore movements being faster than the rate of energy transfer. There the rate of energy transfer has to be calculated taking into account the average over all possible distances and orientations. (b) Intermediate case, called the isotropic average, where the orientational variation of the fluorophores is faster than the rate of energy transfer while the positional variation is slower (c) Static case, where the fluorophore movements are much slower than the rate of energy transfer. In this case each distance and respective fluorophore orientation has to be taken into account with its individual transfer efficiency. These efficiencies then are averaged by the measurement process. (Figure from ref. ²). 2 Wozniak, A. K., Schröder, G. F., Grubmüller, H., Seidel, C. A. M. & Oesterhelt, F. Single-Molecule FRET Measures Bends and Kinks in DNA. Proc. Natl Acad. Sci. USA 105, 18337-18342 (2008).