Fig. 6. De novo assembly of CENPs at the integrated 10 kb alphoid YAC DNA site without chromosome rearrangement. (A-G) Metaphase chromosomes from the del.22HT1-3 cell line, untreated (A,C,E) or treated (B,D,F,G) with 1000 ng/ml of TSA, analyzed by immunofluorescence using antibodies against CENPs (green), and FISH using the YAC arm probe (red). The chromosomes were counterstained with DAPI (blue). Arrowheads indicate the integration sites of the 10 kb alphoid YAC. Scale bars: 2 µm. (H-M) Intensity plot of CENP signals (black) and YAC signals (red) at the 10 kb alphoid YAC site on the host chromosome DNA. (N) The plots of relative intensity of CENP-A (upper), -B (middle), -C (lower) signals at each alphoid YAC integration site on the host centromeres in the del.22HT1-3 cell line untreated (square) or treated (diamond) with 1000 ng/ml of TSA. Red dotted lines indicate the lower limit of signals distinguishable from background levels.

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