Dysbindin knockdown decreases endogenous ciliary polycystin-2 levels. (A) Immunoblot and quantification of total protein levels of MEK cells expressing either pallidin or dysbindin lenti-shRNAs. (Aa) Selected immunoblot images showing pallidin knockdown and (Ab) dysbindin knockdown with either γ tubulin or IFT27 loading control. (Ac) Mean quantification of pallidin and (Ad) dysbindin knockdown under each condition (control, NS-shRNA, shRNA#1, and shRNA#2). Intensity was normalized to their corresponding γ tubulin or IFT27 loading control. shRNA-mediated knockdown of either pallidin or dysbindin results in >90% reduction of total protein abundance. (B) Immunostaining and quantification of ciliary polycystin-2 and cilia length in MEK cells expressing either pallidin or dysbindin lenti-shRNAs. (Ba) Selected images of polycystin-2 (Pkd2) antibody staining (red), Arl13b antibody staining (green), and nuclei are detected with DAPI (blue). Bars, 10 µm. Insets are 170% enlargements of the cilia. (Bb) Mean steady-state ciliary polycystin-2 is decreased and (Bc) mean ciliary polycystin-2 per micrometer is reduced in cells expressing dysbindin lenti-shRNAs. (Bd) Mean cilia length is reduced in cells expressing dysbindin shRNAs. n = 107 cilia per experimental group. Error bars are SEM. Data were analyzed using one-way ANOVA and the Bonferroni multiple-comparisons test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.