Ectopic localization of NEKL-3 inhibits OSM-3 motility. (A) Schematic of plasmids used to ectopically express fused NEKL-3 at transition zone (mksr-2::nekl-3) or distal segment (pcrg-1::nekl-3) under the control of Pdyf-1. (B) Amphid and phasmid cilia in mskr-2::nekl-3 animals were labeled by OSM-3::GFP. The distribution of OSM-3::GFP was not affected. Arrows indicate the ciliary base, and arrowheads indicate the junction between the middle segment and the distal segment. Scale bar, 5 μm. (C) Representative fluorescence images of amphid and phasmid cilia in pcrg-1::nekl-3 animals. Arrows indicate the ciliary base, and arrowheads indicate the junction between the middle segment and the distal segment. Scale bar, 5 μm. (D) Quantification of the cilium length (mean ± SD) measured in the indicated animals. Comparisons were performed between WT and the indicated genotypes by unpaired Student’s t tests. N = 37–48 animals. Statistical significance, compared with the control with a matching color code, is based on Student’s t test. **P < 0.01; ***P < 0.001. (E) Histogram of OSM-3::GFP anterograde IFT velocities along the middle segments (Antero. m.s.). Velocities and numbers of IFT particles (N) are indicated. Each plot was fitted by a Gaussian distribution. Statistical significance, compared with the control with a matching color code, is based on Student’s t test. ***P < 0.001. Data are mean ± SD.