Figure 3

CDK5RAP2 in immortalized lymphocytes and dispersion of centrosomal protein γ-tubulin in CDK5RAP2 mutant patients cells. CDK5RAP2 protein levels were below detection level in immortalized lymphocytes from both patients with the c.4441C > T CDK5RAP2 mutation when assessed by (A) Western blot and (B) immunocytology. Subcellular localization of CDK5RAP2 and γ-tubulin throughout the cell cycle in immortalized lymphocytes of (C) controls and (D) MCPH3 patient 2. In controls, centrosomal CDK5RAP2 levels were weak during interphase, increased in the subsequent prophase and remained high throughout mitosis until telophase, when signals dropped to interphase levels. In the patient cells the alignment of the chromosomes at the spindle poles was less precise than in the control cells. Cells were stained with CDK5RAP2 (red), γ-tubulin (green) as a centrosomal marker, and DNA is stained with DAPI (blue). Scale bars 10 μm. Western blot results reveal that total γ-tubulin protein levels are similar in patients and controls (Additional file 1: Figure S1).