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. 2017 Feb 17;8:14432. doi: 10.1038/ncomms14432

Figure 5. CX-5461 and CX-3543 stabilize G4 sequences.

Figure 5

(a) In vitro FRET melting assay with three different G4 forming DNA fragments and a non-G4 forming dsDNA control. Vertical axis, changes in melting temperature; horizontal axis, drug concentration (μM). Error bars denote the s.d.; n=3. The solid lines represent the interpolation of the values with a single binding curve model. (b) Progression of DNA polymerase was stalled by CX-5461 and CX-3543 when incubating with G4 forming sequence cKit1. Full gel image is displayed in Supplementary Fig. 6c. (c) CX-5461 and CX-3543 bind to and stabilize G4 structure as demonstrated by the increased number of immunofluorescence foci with G4 binding antibody, BG4. Scale bar, 10 μM. Right panel shows the quantification. Median BG4 foci per nucleus is shown. The box extends from the 25th to 75th percentiles. (d) Co-localization between 53BP1 foci and BG4 foci. Drug treatment time is 24 h, N=2.∼500 cells per condition were counted. Scale bar, 10 μM. Right panel shows the quantification. Error bars denote the s.d.