PLP – KHC interaction mutants show reduced centriole motility. (A) Schematic showing the interactions between PLP and KHC and the corresponding interaction mutations (red text). (B) Example projections of PCs showing PLP transgenes (green) localizing centrioles (Asterless; magenta). DNA (blue) shows cells are in interphase. (C) 10-min time projections of centriole movement (colored tracks) in PCs expressing the indicated PLP transgenes in a mutant background (plp²¹⁷²/Df(3L)Brd15). (D) Rescue with the mutant transgenes results in a significantly slower instantaneous velocity compared to full length rescue (PLP::GFP: 117 ± 32, n = 66. PLP^Δ740-971::GFP: 71 ± 25, n = 83. PLP^PD::GFP: 55 ± 32, n = 138, PLP^Δ2539-2895::GFP: 83 ± 36 n = 139, ANOVA: ****P = <0.0001, Dunnett’s pairwise comparison between PLP::GFP and all other conditions). Data = mean ± SD. (E) Mean squared displacement shows motility is most effected by mutations (PLP^PD and PLP^Δ740-971) that interfere with interaction between PLP and the cargo binding tail of KHC. (F) Maximum intensity projections from live NBs showing the position of the MTOCs (yellow arrows; Jupiter::RFP) in prophase. Cells are expressing the indicated PLP transgene in a PLP mutant background (plp²¹⁷²/Df(3L)Brd15). (G) The angle of the centrosomes relative to the nucleus in prophase is not significantly rescued by the transgenes that block PLP–KHC interaction (Control: 139° ± 41.4, n = 25. No Transgene: 72° ± 52, n = 26. PLP::GFP: 134° ± 28, n = 19, P = <0.0001. PLP^Δ740-971::GFP: 54° ± 32.7, n = 18, P = 0.69. PLP^PD::GFP: 85.8° ± 37, n = 27, P = 0.86. PLP^Δ2539-2895::GFP: 104 ± 47.5, n = 25, P = 0.08. P values determined by Tukey pairwise comparison between no transgene and PLP rescue conditions). Data = mean ± SD. (H) Maximum projections showing the localization of the indicated transgenes in interphase NBs. Scale bars: 5 µm. Time stamp: mm:ss.