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. 2015 Jan 8;96(1):81–92. doi: 10.1016/j.ajhg.2014.12.002

Figure 3.

Figure 3

Lack of Dcdc2 Is Rescued by Full-Length DCDC2 and by Wnt Inhibition, but Not by Mutants Detected in NPHP-RC

(A) siRNA (siDcdc2-11) mediated knockdown of Dcdc2 in IMCD3 cells grown in 3D spheroid culture causes a ciliogenesis defect that is rescued by overexpression of human WT DCDC2, but not by mutants found in humans with NPHP-RC. For each condition >50 spheroids were evaluated for the percentage of ciliated cells and the experiment was repeated three times independently. Note that the red structures visible in the mutant-transfected spheroid are midbodies of dividing cells, not cilia. Scale bars are 5 μm.

(B) Ratio of ciliated cells per nuclei within each spheroid upon knockdown and after attempted rescue with WT DCDC2 and two mutants. ∗p < 0.001, as determined by ANOVA analysis. Experiments were repeated at least two times independently and the data combined. Graphs show mean value and standard error of the mean (SEM).

(C) The ciliogenesis defect was rescued by growing spheroids in medium treated with Wnt inhibitor iCRT14 (100 μM). Scale bars are 5 μm.

(D) Quantification for two siRNAs of rescue of ciliogenesis defect by iCRT14 treatment. ∗p < 0.001, as determined by ANOVA analysis. Experiments were repeated at least two times independently and the data combined. Graphs show mean value and SEM.