Research Paper Volume 8, Issue 11 pp 2827—2847

Figure 2. CDC13-EST2 expression enhances genome instability. (A) ERC level in fob1∆ and sir2∆ mutant cells was examined by Southern blot with a probe of 25S rDNA sequence (upper panel). One species of multimer ERCs (indicated by solid arrow) was quantified and normalized to genomic rDNA signal. Lower panel is the quantification of the ERC level in fob1∆ and sir2∆ mutant cells. The ERC level in wild-type cells was set as “1”, and the ERC value in each strain was normalized to the genomic rDNA. (B) and (C) Detection of ERC level in young cells of CLS D1 (B) and old cells of CLS D27 (C) used in Fig. 1E by Southern blot (upper panel). One species of multimer ERC (indicated by solid arrow) was quantified (lower panel). POL1 gene was used as an internal loading control. Values in the quantification are normalized to POL1 level ± SEM. (D) Chronological viability of cells carrying CAN1-URA3 cassette at CLS D1, D15 and D29. The cells of streakout 8^th containing pRS315 (control) and pRS315-CDC13-EST2 was examined. Survival (viable colonies) values were normalized to CLS D1. Values are the averages of 6-10 cultures ± SEM. (E) and (F) CAN1 mutation frequency (E) and GCRs frequency (F) in overlong- and normal-telomere cells at CLS D1, D15 and D29 were examined. Values in (E) and (F) are the averages of 6-10 cultures ±SEM. (G) Telomere length analysis of cells at CLS D1 and D29. The cells of streakout 8^th that containing pRS315 (normal telomere) and pRS315-CDC13-EST2 (overlong telomere) were examined.