Fig. 6: Use of Deep-LASI on titration experiments, protein data, and confocal data.

a, b 3cFRET: Tuning the dissociation thermodynamics between protruding ssDNA strands by osmolytes. a CDFs of the dwell times, assessed through mono-exponential fits, for state 1 (left) and state 2 (right) of the L-shaped DNA origami structure from Fig. 4a decrease with increasing glycerol concentration. b Dependence of dwell times for both states versus glycerol concentration. c–f 2cFRET: Probing domain-domain interactions in Ssc1, a mitochondrial Hsp70. c Frame-wise smFRET distributions of Hsp70 molecules in the presence of 1 mM ADP classified as dynamic by Deep-LASI (gray) and evaluated manually (blue) from a total of 3534 traces. d The TDP generated by Deep-LASI aligns with the data plotted in (c), illustrating the interconversion between the undocked ( ~ 0.5) and docked ( ~ 0.8) conformations. Total number of transitions, n = 3914. e CDFs of the dwell times and mono-exponential fits to the dwell time distributions derived by Deep-LASI for domain docking (left panel) and domain undocking (right panel) depending on the ADP concentration. f Comparison between average dwell times extracted by Deep-LASI (triangles) and by manual evaluation (crosses) using HMM. Deep-LASI matches the published trend with similar dwell times⁴⁰. g–i 1c-FRET: Deep-LASI analysis of ssDNA binding kinetics observed via confocal microscopy. g Confocal trace (with 2 ms binning) of the DNA origami structure from Fig. 2a with 6 nt binding sites and corresponding states predicted with high confidence. h Confocal trace (with 0.6 ms binning) of a DNA origami structure with 5 nt binding sites and predicted states. Due to the low SNR of the data, the confidence output of Deep-LASI reaches its lower limit. i Mean dwell times obtained from confocal data for various binding site lengths analyzed by sg-FCS⁴¹ (circles) and Deep-LASI (triangles). The results align well, except for dwell times extracted from the 5 nt sample, which was predicted with a low confidence distribution due to low SNR and a limited amount of information in the one-channel input. Dwell times obtained from TIRF data are displayed in light gray for comparison. nt: nucleotides. Source data are provided as a Source Data file.