Fig. 3: Dynamics of single OSM-3 (OSM-3::mCherry), IFT dynein (XBX-1:eGFP) and tubulin (TBB-4::eGFP) molecules entering cilia.

a Distance-time plots of OSM-3 (419 tracks from 16 cilia). b Distribution (upper panel) and histogram (lower panel) of docking locations of OSM-3 (average docking location is 334 ± 39 nm). The distribution of docking locations of kinesin-II is overlayed (green line). c Distribution of the measured pause time, \({t}_{{p\_m}}\), with respect to actual pause time, \({t}_{{p\_actual}}\), for OSM-3, obtained from numerical simulations (using \({t}_{{bleach}}\) 1.4 s). Each point represents the average \({t}_{{p\_m}}\) for a single simulated experiment of 1500 events (example simulation in Supplementary Fig. 3c), varying \({t}_{{p\_actual}}\) by steps of 0.01 s. Simulated experiments yielding \({t}_{{p\_m}}\) 0.4 ± 0.1 s (similar to experimental value; Supplementary Fig. 4a) are highlighted in red, indicating that \({t}_{{p\_actual}}\) is in the range 0–0.1 s. d Distance-time plots of IFT dynein (666 tracks from 14 cilia). e Distribution (upper panel) and histogram (lower panel) of docking locations of IFT dynein (average docking location is 112 ± 8 nm). f Distribution of the measured pause times, \({t}_{{p\_m}}\), with respect to actual pause times, \({t}_{{p\_actual}}\), for IFT dynein, obtained from numerical simulations (using \({t}_{{bleach}}\) 1.8 s). Each point represents the average \({t}_{{p\_m}}\) for a single simulated experiment of 10,000 events (example simulation in Supplementary Fig. 3d), varying \({t}_{{p\_actual}}\) by steps of 0.01 s. Simulated experiments yielding \({t}_{{p\_m}}\) 1.8 ± 0.2 s (similar to experimental value; Supplementary Fig. 4b) are highlighted in red, indicating that \({t}_{{p\_actual}}\) is in the range 2.7–8 s. g Distance-time plots of tubulin (170 tracks from 11 cilia). h Distribution (upper panel) and histogram (lower panel) of docking locations of tubulin (average docking location is 660 ± 255 nm). i Distribution of binned average velocities (solid line) along the length of the cilium for kinesin-II (green), OSM-3 (orange), IFT dynein (purple), tubulin (blue), with the shaded areas indicating the errors. Average values and errors are estimated using bootstrapping (see “Methods”).