Overexpression of MyoVb perturbs polycytin-2 trafficking to the primary cilium. (A) Selected trafficking images of polycystin-2-GFP-SNAP IMCD Flp-In cells overexpressing Flag-MyoVb C-terminal tail. GFP and Arl13b antibody staining (green), SNAP TMR STAR (red), and Flag-MyoVb (blue). Insets depict newly synthesized polycystin-2-GFP-SNAP trafficking to the cilium in control or Flag-MyoVb-overexpressing cells. Insets also show accumulation of newly synthesized polycystin-2-GFP-SNAP in the recycling endosome in the Flag-MyoVb-overexpressing cells. Insets are 200% enlargements of the cilia and 170% enlargements of the recycling endosome. Bars, 10 µm. (B) Mean cilia length of polycystin-2-GFP-SNAP IMCD Flp-In control and Flag-MyoVb-overexpressing cells after the Golgi release. Cilia are shorter in Flag-MyoVb-overexpressing cells. (C) Mean SNAP ciliary fluorescence of Polycystin-2-GFP-SNAP delivery to the cilium after Golgi release. Polycystin-2 ciliary trafficking decreased in Flag-MyoVb-overexpressing cells. (D) Linear regression slope values of newly synthesized polycystin-2-GFP-SNAP in IMCD Flp-In control and Flag-MyoVb-overexpressing cells after the Golgi release using data points taken at 0, 1, 2, 4, and 6 h. Mean SNAP ciliary fluorescence and mean ciliary length were plotted from three independent experiments in which 30 cilia were quantified for each condition (control and Flag-MyoVb) at each time point (n = 90 total cilia per time point). Error bars represent SEM. Data were analyzed using the unpaired Student’s t test. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.