Figure 3.

Biased Mis-segregation of Chromosomes 1 and 2 Is Not Dependent on Kinetochore Expansion

(A) Immunofluorescence images of RPE1 cells treated with monastrol or nocodazole for 8 hr as indicated, stained with antibodies to mark centromeres (CREST serum, red) and outer kinetochores (CENP-E, green).

(B) Kinetochore size quantification.

(C) RPE1 cells were treated with 8 hr monastrol, then released for 1.5 hr before FISH with specific centromere enumeration probes as indicated.

(D) Segregation error rates and average number of lagging chromosomes per erroneous anaphase.

(E) Quantification of percentage of lagging chromatids that are the chromosome indicated from erroneous anaphases (77–299 lagging chromosomes analyzed per chromosome).

(F) Immunofluorescence-FISH images of cells treated with nocodazole for 8 hr and stained with CREST sera, anti-CENP-E, and FISH using CEP1.

(G and H) Quantification of outer kinetochore intensity (G) or expanded kinetochore size (H) at chromosome 1 compared with other chromosomes after 8 hr nocodazole.

All experiments show mean ± SD of at least three independent experiments. ^∗∗p < 0.005 and ^∗∗∗∗p < 0.00005. See also Figure S5.