Complementary results for GFP-PBP1b and GFP-RodZ tracks. (a) Results from parameter fits to experimental tracks of GFP-PBP1b of at least three time points (and considering not more than the first 50 time points) assuming two, three, or four states in ExTrack (three replicates per condition, each replicate has at least 17,000 tracks of average lifetime from 6.3 to 7.5 positions). ExTrack settings: Window length = 4 for two/three states and 5 for four states, no sub-steps. State fractions obtained from rates. (b–d) ExTrack analysis of GFP-RodZ data (same as in Fig. 4, f–h). (b) Parameters found by ExTrack for GFP-RodZ tracks (three replicates, each replicate has at least 25,000 tracks of average lifetime from 6.3 to 7.4 positions), considering all tracks with at least three time points and restricting analysis to the 50 first time points. (c) Histogram of the time spent in bound or diffusive states, among tracks of at least 21 time points using parameters from b. (d) Examples of long bound RodZ tracks in linear motion. (e) PBP1b rebinding. Histograms of the distances in between initially bound position and after four diffusive steps of PBP1b tracks that are subsequently either rebinding (blue) or not rebinding (red; see Materials and methods). Histograms show no noticeable difference. Error bars and shaded regions: SDs between replicates.