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. 2018 Dec 4;19(12):3865. doi: 10.3390/ijms19123865

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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smFRET reveals Sox2^HMG concentration-dependent multistep bending of DNA^NANOG. (a) E_FRET histograms of DNA^NANOG with increasing [Sox2^HMG]. (b) [Sox2^HMG]-E_FRET contour map color coded based on fractional occupancy of individual DNA conformations. Corresponding DNA conformations are marked on the contour map. (c) Sox2 binding isotherm of the U ⇋ B_I transition as probed by detecting changes in E_FRET, linked to dsDNA bending transition. The NLS-derived apparent K_D for this binding step is 0.30 (±0.04) nM (binding equation with fixed Hill coefficient of 1). (d) dsDNA^NANOG conformational distributions as modulated by Sox2^HMG concentration, determined from NLS fitting of individual smFRET histograms to Gaussian functions.