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. Author manuscript; available in PMC: 2016 Dec 17.
Published in final edited form as: Mol Cell. 2015 Nov 19;60(6):886–898. doi: 10.1016/j.molcel.2015.10.027

Figure 1. Inducible CRISPR knockouts define the contributions of the CCAN to chromosome segregation.

Figure 1

A) Diagram of the CCAN components in their sub-complexes. B) Strategy for the generation of inducible knockouts of CCAN genes. spCas9 under control of a tet-on promoter is stably integrated into HeLa cells and sgRNAs targeting early exons of the gene of interest are stably integrated by lentiviral transduction. C) Representative immunofluorescence images of mitotic phenotypes following CENP-N inducible knockout. D) Quantification of mitotic phenotypes following inducible knockout of a component of each CCAN subcomplex for four days. Cells were classified as having an alignment defect if they displayed bipolar-like spindles with > 4 off-axis chromosomes. n = 100 cells per condition. E) Representative immunofluorescence images of interphase phenotypes following CENP-C inducible knockout. F) Quantification of interphase phenotypes following inducible knockout of a component of each CCAN subcomplex for four days. n = 100 cells per condition. Scale bars, 5 μm.