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. 2003 Sep;14(9):3821–3833. doi: 10.1091/mbc.E03-01-0860

Figure 8.

Figure 8.

Chromosome segregation defects in TSA-treated MRC-5 cells derive from defective sister chromatid separation. (A) Immunofluorescence detection of kinetochores and DNA staining on anaphase cells, showing lagging chromosomes or chromatin bridges. The lagging chromosome shows two paired CREST signals (arrow). The chromatin bridge shows two extended CREST signals at the ends of the bridge (arrows). Bar, 5 μm. (B) Quantitative analysis of the induction of chromosome segregation defects after TSA treatment. Lagging chromosomes were classified as showing one CREST signal (single sisters) or two paired CREST signals (paired sisters) both in DMSO- and TSA-treated cells. Chromatin bridges, both in TSA-treated cells and DMSO-treated cells, were classified as follows: without identifiable CREST signal (without CREST signal); with CREST signal at opposite ends of the bridge (CREST distal); with CREST signal along the chromatin bridge (CREST central). The graph shows results obtained scoring 937 anaphase cells in DMSO-treated cells and 921 anaphase cells in TSA-treated cultures. Data from 1-h and 7-h treatments with TSA were pooled because no appreciable differences were observed. (C) Fluorescence in situ hybridization with chromosome 16-specific alphoid probe (fluorescein isothiocyanate, green) and chromosome 1 classical satellite probe (rhodamine, red) on anaphase MRC-5 cells after treatment with TSA. DNA was counterstained with DAPI (blue). A regular distribution of the two chromosomes is expected to give two green signals and two red signals on each group of segregated chromosomes (normal cell). Some chromatin bridges on anaphase cells showed two extended red (chromatin 1 bridge) or green (chromosome 16 bridge) signals at the opposite ends of the connecting chromatin. In other cases, the centromeric signal totally overlapped the chromatin bridge (centromeric bridge). Anaphases (519) were scored in TSA-treated cells and 11 chromatin bridges involving either chromosome 1 or chromosome 16 were recorded. Two of 11 were centromeric bridges.