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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: Aging Cell. 2013 May 15;12(4):635–644. doi: 10.1111/acel.12085

Figure 3. Telomerase is required for telomere lengthening in the tsa1 mutant.

Figure 3

(A) The wild-type and tsa1 spores from a TSA1/tsa1 heterozygous strain were re-streaked successively on YEPD plates. Genomic DNA was isolated at the indicated time and analyzed for telomere length using Southern blot as in Figure 1A. (B) Telomere XVL from either wild-type or tsa1 cells was amplified using a high fidelity DNA polymerase, cloned, and sequenced. Telomere sequences were aligned and constant and divergent sequences were indicated by blue and red color. The average length of constant and divergent sequences is shown below the graph. Error bars indicate standard errors. NS, not significant; *, p<0.05. (C) Telomeres were categorized according to the length of constant sequences and the length of divergent sequences and compared between the wild-type and tsa1 cells. Error bars indicate standard errors. NS, not significant; *, p<0.05. (D) Spores with the indicated genotype generated from a TSA1/tsa1 EST2/est2 heterozygous strain were re-streaked successively on YEPD plates for ~ 90 generations. Genomic DNA was analyzed for telomere length using Southern blot as in Figure 1A. Representative clones are shown. Calculated Y′-telomere lengths (mean±SD) from three clones for each genotype are shown at the bottom.