Genome-wide analysis of ectopic CENP-A demonstrates enrichment of CENP-A hotspots at DNase I hypersensitive sites. (A) Concordance analysis of chromatin immunoprecipitation (ChIP)-seq replicates of ectopic CENP-A IPs in normal colon, HeLa, and SW480 cells displays r2 > 0.9, indicative of high reproducibility. (B) Left: distribution analysis of hotspots in normal, HeLa and SW480 cells; Venn diagrams of hotspots from SW480, HeLa, or normal colon cells versus Mock immunoprecipitation (IP). Right: Genomic location of hotspots from left panel shown as a percentage of total. Legend below indicates genomic location. Fisher’s exact test was performed for comparison of HeLa and SW480 histograms: P = 0.0174 (C) Genome browser snapshots of ectopic CENP-A localization depict representative examples of CENP-A and DHS hotspots at genes overlapping with clusters of DNase I sensitivity, H3K27Acetylation and Transcription factor binding in the ENCODE data. Scale bars are in the upper left corner of each snapshot. (D) Distribution of HJURP hotspots in HeLa and SW480 cells. Venn diagram demonstrates 30 to 40% of HJURP hotspots overlap with ectopic CENP-A hotspots, but the number of HJURP hotspots in SW480 is very low.