Journal: Nature Communications

Article Title: Structural role for DNA Ligase IV in promoting the fidelity of non-homologous end joining

doi: 10.1038/s41467-024-45553-z

Figure Lengend Snippet: a Representative single-molecule trajectories showing Lig4 colocalization, Lig4 end binding, and synapsis. Panels as in Fig. . b For molecules that underwent SR synapsis, Cy7 colocalization trajectories (e.g., bottom of panel a ; n = 248 from 8 independent experiments) were aligned, with the onset of SR synapsis corresponding to t = 0 (gray bar). Blue line shows median Lig4 stoichiometry as a function of time to synapsis. See Supplementary Data Fig. for distributions. c Scatter plot depicting Cy3B/Cy7 FRET and Cy5/Cy7 FRET at the moment of SR synapsis, as detected by a stepwise increase in Cy3B/Cy5 FRET using the MATLAB ischange function. Only synapsis events for which colocalization of Cy7-labeled Lig4 was detected are shown. Likely due to incomplete Cy7 labeling, Lig4 colocalization was detected in 178 of 248 total synapsis events. n = 178 from 8 independent experiments.

Article Snippet: To convert Cy7-Lig4 intensities to stoichiometries, stepwise changes (corresponding to Lig4 binding/unbinding events) were detected in the Cy7 ex./Cy7 em. trajectory for each DNA molecule using the MATLAB ischange function.

Techniques: Binding Assay, Labeling