Figure 4

Ablation of RAD50, NPM1 and XRCC5 lead to genomic instability and facilitate multinucleated cells and mitotic irregularities. A. Representative Images of control siRNA, siNPM1, siRAD50 and XRCC5 A4-T cells - panels indicate Giemsa (top), Hoechst (middle) and α-tubulin stained (lower) nuclei respectively. Marked arrow indicating incidences of abnormal mitotic figures, multi-nucleation and mitotic irregularities in the siRAD50, siNPM1 and siXRCC5 cells respectively, while in control siRNA transfected cells it is denoting normal mitosis; 40X magnification; 50 μm scale bar. Representative graphs indicating frequencies of mitosis and abnormal mitotic figures in control and siRAD50, siNPM1 and siXRCC5 cells. B. Representative images of cells stained for α- and γ- tubulin expression indicating existence of abnormal spindles as well as numbers and position of centrosomes during mitosis in control and siRAD50, siNPM1 and siXRCC5 cells; representative graphs indicating frequencies of aberrant spindle formation and cells with more than 2 centrosomes in control and siRAD50, siNPM1 and siXRCC5 cells on the right. C. Graphical representation of flow cytometry based resolution of cell cycle phases and aneuploid genomic content associated with indicative of control and siRAD50, siNPM1 and siXRCC5 cells through PI staining. Data shown as mean ± SE of triplicate experiments. *p <0.05; **p <0.01.