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. 2015 Nov 17;43(22):10907–10924. doi: 10.1093/nar/gkv1225

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — SsoSSB binding to a short ssDNA segment leads to single-molecule trajectories showing coexisting FRET, PIFE and acceptor quenching events. (A) Representative single-molecule trajectory showing the effect that SsoSSB binding to a surface-immobilized 12-mer ssDNA labelled at the 3′ end with Cy3 (BidC₁₂Cy3) has on the donor and acceptor intensities (upper panel), the PIFE signal (middle panel) and the FRET efficiency (bottom panel). Panels on the right show the single-molecule histograms for the corresponding trajectory displayed on the left. (B) Single-molecule trajectories and corresponding histograms obtained at identical conditions as those described for (A) but showing additional rare events (marked with an asterisk). As shown in panel (A), the majority of PIFE events formed quickly following a high-FRET state and only a small percentage (panel B, marked with asterisk) was observed to directly emerge from a non-FRET state (no protein bound).