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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Nat Cell Biol. 2013 Nov 24;15(12):1495–1506. doi: 10.1038/ncb2879

Figure 4.

Figure 4

Hypomethylation and expression of late-replicating satellite sequences in senescent cells. (a) Bisulfite sequencing of satellite 2 in proliferating and senescent cells. Each row represents an individual clone. Filled and open circles are methylated CpG and unmethylated CpG dinucleotides respectively. (b) Southern blot of genomic DNA with the methyl-sensitive enzyme BstBI probed with a probe specific for satellite 2. Genomic DNA was harvested at different timepoints as cells approach senescence as indicated by arrow. (c) FISH using a probe for satellite 2 in proliferating and near-senescent cells. Scale bar, 5µm. (d) Measurement of two-dimensional (2D) area occupied by the FISH probe from c. (e) RT-PCR of satellite 2 in IMR90 cells at indicated PD and senescence. (f) FISH using a probe for MYC in proliferating and near-senescent cells as a negative control for c. Scale bar, 5µm. (g) Measurement of the two-dimensional area occupied by the MYC FISH probe in f. For all graphs, error bars indicate the mean ± s.e.m. of three independent experiments where 100 cells were scored per experiment. Source data for d,g in Supplementary Table 22.