Kinesin-1 is required for efficient centriole motility. (A) 10-min time projections of centriole movement (colored tracks) in the indicated knockdown conditions in PCs. Scale bars: 5 µm. (B) Average velocity is significantly reduced following the knockdown of Kinesin-1 components (control: 112 ± 20, n = 3 wing discs, 100 centrioles. KHCRNAi: 71 ± 5, n = 3 wing discs, 110 centrioles. KLCRNAi: 92 ± 5, n = 4 wing discs, 97 centrioles. EnsRNAi: 83 ± 8, n = 6 wing discs, 84 centrioles; data = mean ± SD. ANOVA P = 0.001, Dunnett’s pairwise comparisons: Ctrl vs. KHCRNAi P = 0.0005, ***, Ctrl vs KLCRNAi P = 0.043, *, Ctrl vs. EnsRNAi P = 0.0027, **). (C) Mean squared displacement is reduced following KHC knockdown in PCs. (D) Diagram summarizing the models by which Kinesin-1 could move centrioles in cells. Kinesin-1 cargo domain is shown in yellow; motor domain (orange) always walks toward the indicated + sign, and the black arrow indicates the movement direction of the centriole. (E) Z-stack projections of fixed NBs sowing centriole positioning. Note: in khc⁸/khc⁶³ NBs, the centrioles are adjacent to the apical side of the cell. (F) Quantification of the percentage of neuroblasts with adjacent apical centrioles. y,w: 17.8% ± 7, n = 5 brains; khc⁸/khc⁶³: 92.5% ± 3, n = 4 brains, khc^mut.A: 17.4% ± 3.5, n = 4 brains. Data = mean ± SD. (G) Averaged OMX-SIM micrograph showing mNG::KHC localizes to the outer centriole edge. Scale bar: 500 nm. (H) Quantification of rotational averaged centrioles showing the distribution of mNG::KHC relative to PLP and RFP::SAS-6 (n = 4).