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. 2017 Mar 7;8:14678. doi: 10.1038/ncomms14678

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Copyright © 2017, The Author(s)

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(a) Schematic of the CENP-A shRNA experiment. (b) Western blot showing endogenous and tagged CENP-A levels following shRNA treatment. (c) CENP-A shRNA mediated suppression causes multipolar spindles in p53^−/− cells, but not in p53^+/+ HCT116 cells. Cells were stained for α-tubulin. (d) Quantitation of the multipolar cells. (e,f) Wild-type and p53 null cells both showed increased lagging chromosomes after CENP-A shRNA mediated suppression. Replacement with methylation mutant CENP-A in p53^+/+ and p53^−/− HCT116 failed to rescue the chromosome segregation defects. Scale bars, 10 μm (g,h) Percentage of cells showing lagging chromosomes after CENP-A knockdown and replacement in the indicated cell lines. The experiments were done in duplicate. Error bars indicate s.d. Indicated P-values were determined by χ² test.