WDR-60 expression is restricted to ciliated sensory neurons, where it has distribution and IFT kinetics similar to those of dynein-2 HC. (A) Endogenously tagged WDR-60::3xFLAG::GFP is expressed in the same ciliated neurons that express dynein-2 HC (GFP::CHE-3). These are the same neurons that incorporate the DiI lipophilic dye. The top illustration shows the relative localization of amphid and phasmid ciliated neurons in C. elegans.(B) Phasmid cilia coexpressing GFP::CHE-3 and XBX-1::RFP. (C) Quantification of GFP::CHE-3 signal intensity along cilia (n = 109 cilia). (D) Phasmid cilia coexpressing WDR-60::3xFLAG::GFP and XBX-1::RFP. (E) Quantification of WDR-60::3xFLAG::GFP signal intensity along cilia (n = 101 cilia). (F) Cilium kymographs of GFP::CHE-3 and WDR-60::3xFLAG::GFP. Single and merge channels for particles moving anterogradely and retrogradely are shown. (G) Mean IFT frequency of anterogradely and retrogradely moving GFP::CHE-3 and WDR-60::3xFLAG::GFP particles per second (n ≥ 22 cilia). (H and I) Anterograde and retrograde velocities of GFP::CHE-3 and WDR-60::3xFLAG::GFP particles along cilia. (J and K) Mean velocities for each cilium subcompartment (n ≥ 430 particle traces were analyzed in ≥ 8 cilia). B, cilium base; TZ, transition zone; MS, middle segment; DS, distal segment. XY velocity and intensity distribution graphs are shown as mean ± SEM, and graphs in columns are shown as mean ± SD. Student’ t test was used to analyze the datasets in G, J, and K. Scale bars: 10 µm (A); 2 µm (B and D); vertical 5 s, horizontal 2 µm (F).