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. Author manuscript; available in PMC: 2012 Jul 12.
Published in final edited form as: Curr Biol. 2011 Jun 30;21(13):1158–1165. doi: 10.1016/j.cub.2011.06.015

Figure 3. Aurora B enrichment at misaligned chromosomes leads to increased phosphorylation of Dsn1 and is a dominant feature of RPE cytoplasm.

Figure 3

(A–D) RPE (panels A and B) and HeLa (panels C and D) cells were subjected to monastrol washout, fixed and stained for immunofluorescence. Representative images are shown in panels A and C and quantification is shown in panels B and D of levels of Dsn1 phosphorylated at Ser100. CENP-C is used as a kinetochore marker. In all cases, the relative intensity of the target at aligned chromosomes is normalized to 1 A.U. The insets are 2X magnified views of the boxed area. Black dashed lines (B and D) indicate the mean of relative intensity for each population.

(E) Scheme for cell fusion experiments starting with HeLa and RPE cells stably expressing CENP-A fusion proteins (HA-tagged in HeLa and YFP-tagged in RPE) that mark the cell line of origin for every chromosome in our analysis.

(F) Quantification of INCENP levels on adjacent co-seeded (but unfused) cells imaged on the same coverslip. Error bars represent standard error of the mean.

(G) Quantification of INCENP levels on misaligned and aligned chromosomes originating from the indicated cell line in fused RPE:HeLa cells. Error bars represent standard error of the mean.

(H) Image of a fused RPE:HeLa cell. The insets are 3X magnified views of the boxed area. Scale bars = 2 µm in panels A, C, and H.