CENP-A OE and mislocalization contribute to CIN, aneuploidy, and karyotypic heterogeneity in cell line and a xenograft mouse model. (A) Observation: Mislocalization of CENP-A contributes to CIN phenotypes, chromosome mis-segregation, and micronuclei formation with ruptured nuclear membrane. (B) Mechanism: In control cells (left) without CENP-A OE and mislocalization, the levels of kinetochore- and centromere-associated proteins are normal, with normal pulling force between two sister kinetochores; however, cells with CENP-A OE and mislocalization (right) show mislocalization of CENP-C, reduced levels of outer kinetochore proteins (CENP-T, NUF2, HEC1, and Mis12), resulting in unstable KT-MT attachments and reduced interkinetochore distance. (C) Physiological consequences: Constitutive OE of CENP-A contributes to aneuploidy and karyotypic heterogeneity in cell line and a xenograft tumor model.