Figure 8: Phosphorylation site mutants of CEP170 and NuMA induce mitotic defects.

(a) HeLa cells were transfected with CEP170 or NuMA siRNA to deplete the endogenous levels of these proteins and subsequently transfected with GFP-tagged wild-type or mutant constructs of CEP170 or NuMA. Cells were fixed and subsequently stained for Alpha tubulin (red). Mutant CEP170 or mutant NuMA transfected cells showed mitotic defects compared to wild-type CEP170 or NuMA transfected cells. (b) Quantification of cells with mitotic defects. Graphs represent average of two experiments. Error bars represent s.d. (c) Kinase assay with wild-type or mutant GST tagged CEP170 or NuMA as substrates. Images shown are representative of three independent experiments. (d) Immunoprecipitation–western blot analysis demonstrating the binding of Kif2b and wild-type or mutant CEP170 (e) Immunoprecipitation–western blot analysis demonstrating the binding of dynein and wild type or mutant NuMA.