Overexpressed CENP-A contributes to increased incidence of micronuclei with ruptured nuclear membrane in DLD1 cells. (A) Increased incidence of micronuclei in DLD1 cells with overexpressed and mislocalized CENP-A. Images show micronuclei in DLD1^CENP-A cells with or without DOX treatment. Cells were immunostained with antibodies against CENP-A, stained with DAPI, and analyzed for the presence of micronuclei. Yellow arrows show micronuclei in DOX-treated DLD1^CENP-A cells. Scale bar: 5 µm. (B) Quantification of images from A shows the incidence of micronuclei in DLD1^CENP-A cells with or without DOX treatment. (C) Micronuclei with different status of nuclear membrane in DLD1 cells with overexpressed and mislocalized CENP-A. Images show micronuclei with normal nuclear membrane, ruptured nuclear membrane, or no nuclear membrane in DLD1^CENP-A cells with DOX treatment. Cells were immunostained with antibodies against CENP-A and Lamin B and stained with DAPI. Insets correspond to the z-stacks from the white-boxed area in the main figure. Scale bar: 5 µm (main image), 1 µm (insets). (D) Quantification shows the proportion of micronuclei with different status of nuclear membrane in DLD1^CENP-A cells with or without DOX treatment. Error bars in B and D represent SEM from three independent experiments. P values were calculated using the unpaired Student’s t test.