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. 2021 Jan 8;12:162. doi: 10.1038/s41467-020-20414-7

Fig. 1. In vitro fluorescence-lifetime of DAOTA-M2 bound to different DNA topologies.

Fig. 1

a Chemical structures of the DNA binders under study in this work. b Time-resolved fluorescence decays of DAOTA-M2 (2 µM, black trace) and following the subsequent additions of dsDNA (CT-DNA, 20 µM, green trace) and then G4 (c-Myc, 4 µM, red trace). Instrument response function (IRF) is shown in grey. Recorded data is shown as dots and fitted biexponential function as a solid line. c Variation of the average lifetime (τw) of DAOTA-M2 in the presence of different G4s (red dots), ss/dsDNA (green dots) and ss/dsRNA (blue dots), adapted from reference32. d Fluorescence-lifetimes of DAOTA-M2 (2 µM) in aqueous buffer (black dot), buffered Xenopus egg extract (33 µL egg extract + 12 µL aqueous buffer, black dot), and in buffered cell extract supplemented with G4 (4 µM c-Myc, red dot) and dsDNA (44 µM ds26, green dot). Both measurements remained constant over 0.5 hr incubation at 21 °C. Mean value from 3 independent measurements, error bars represent standard deviation. e In a mixture of DAOTA-M2 (2 µM), dsDNA (CT-DNA, 20 µM), and G4 (c-Myc, 4 µM), increasing amounts of PDS (5.3. 7.9. 9.9, and 14.4 µM, black traces) displaces DAOTA-M2 from a G4 to dsDNA environment. Recorded data is shown as dots and fitted biexponential function as a solid line. f Variation of τw in a mixture of DAOTA-M2 (2 µM), dsDNA (CT-DNA, 20 µM), and G4 (c-Myc, 4 µM), with increasing concentrations of G4 binders PDS (black dots) and Ni-salphen (orange dots), and a non-G4 binder DAPI (purple dots). See Figure S2 for example decays. Mean value from 3 independent measurements, error bars represent standard deviation. g Variation of τw in a mixture of DAOTA-M2 (2 µM) and G4 (c-Myc, 4 µM), with increasing concentrations of Zn (grey dots), VO (purple dots), Ni (orange dots), and Cu (green dots)-salphens. Mean value from 3 independent measurements, error bars represent standard deviation. Unless stated otherwise, all experiments in 10 mM lithium cacodylate buffer (pH 7.3) with 100 mM KCl. Source Data are available as a Source Data file for Fig. 1b–g.