Fig. 6

CENP-B and Daxx knockout reduce levels of ATRX and H3K9me3 at centromeres. Top: Representative images of MCF7 control (left column), CENP-B knockout (middle column), Daxx knockout (right column). Top row: ATRX (green) is associated with subpopulation of centromeres (CENP-A, red) and ND10/PML bodies (white arrow in inset) in MCF7 control cells (left); the association of ATRX with centromeres is reduced in CENP-B (middle) and Daxx (right) knockout cells. Note that there is no ND10-like ATRX staining in Daxx knockout cells since Daxx is required for ATRX recruitment to ND10/PML bodies [35]. Bottom row: CENP-B (middle) and Daxx (right) knockout reduce levels of H3K9me3 at centromeres when compared to control cells (left). Graphs in the bottom panel show fluorescence intensity along the region of interest (ROI). Bottom: Quantitative analysis of intensity of ATRX and H3K9me3 staining at centromeres. Data are shown relative to wild-type cells (100%). Statistical analysis was performed by one-way ANOVA followed by Tukey–Kramer multiple comparison test. The bars represent mean ± SD; > 200 cells from 10 random fields were analyzed for every clone