Figure 5
Terminal restriction fragment analysis coupled with Southern blot was used for telomere length analysis. The stable over-expression of wild-type PML and PML C/C- did not affect telomere length in U2OS cells. A) (i) Southern blot analyses of telomere length in U2OS PCI Neo STC6, PML STC20 and PML C/C- STC20. (ii) Graphical representation of telomere length of clones in (A) (i). B) (i) Southern blot analyses of telomere length in U2OS PCI Neo STC8, PML STC21 and PML C/C- STC22. (ii) Graphical representation of telomere length of clones in (B) (i). Telomere length changes were observed in MCF7 cells following stable expression of wild-type PML and PML C/C-. C) (i) Southern blot analyses of telomere length in MCF7 PCI Neo STC1, PML STC7 and PML C/C- STC7. (ii) Graphical representation of telomere length of clones in (C) (i). D) Southern blot analyses of MCF7 PCI Neo STC4, PML STC10 and PML C/C- STC22. (ii) Graphical representation of telomere length of clones in (D) (i). Genomic DNA was obtained from the cells at every 5th passage until the 20th passage. Genomic DNA was digested with HinfI and RsaI and the digested DNA was resolved on an agarose gel. Southern blot analysis was carried out using the TeloTAGGG telomere length assay kit according to manufacturer’s protocol (Roche). The smears represent the telomeres detected and grey lines indicate the mean telomere length as analysed with Kodak Imager.