The Rate of Increase of Short Telomeres Predicts Longevity in Mammals Elsa Vera, Bruno Bernardes de Jesus, Miguel Foronda, Juana M. Flores, Maria A. Blasco  Cell Reports  Volume 2, Issue 4, Pages 732-737 (October 2012) DOI: 10.1016/j.celrep.2012.08.023 Copyright © 2012 The Authors Terms and Conditions

Figure 1 Mouse Telomeres Shorten 100-Fold Faster than Human Telomeres with Aging (A) Kaplan-Meyer survival curves of the indicated cohorts. Arrows indicate time of blood extraction. (B and C) The mean telomere length (B) and percentage of short telomeres (C) were determined by HT-QFISH on white blood cells (n, number of mice). Values are given as average ± SEM, and statistical significance was determined by one-tailed Student’s t test. (B) From left to right (average ± SEM): 51.49 ± 1.633, 45.10 ± 1.322, 43.18 ± 1.118, 37.51 ± 2.241, 52.61 ± 2.049, 46.06 ± 2.142, 40.54 ± 2.053, 38.55 ± 1.801. (C) From left to right (average ± SEM): 12.78 ± 0.8693, 16.19 ± 0.8230, 17.99 ± 1.257, 25.14 ± 3.552, 12.16 ± 1.078, 16.68 ± 1.237, 20.77 ± 1.798, 21.93 ± 2.581. (D and E) Adjustment of mean telomere length (D) and percentage of short telomeres (E) with aging to a linear or quadratic model. Second-order polynomial adjustment (quadratic) was used for the nonlinear fit model. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure 2 The Rate of Increase in the Percentage of Short Telomeres and the Rate of Telomere Shortening Predicts Mouse Longevity (A–C) The percentage of short telomeres (<15 kb; A and C) and mean telomere length (B and C) were determined by HT-QFISH on white blood cells (see Figure 1). Linear regression lines were calculated for the values obtained for the rate of increase in the percentage of short telomeres and for the rate of mean telomere length shortening (kb/month) versus lifespan (weeks). (A) Rate of increase in the percentage of short telomeres (<15 kb) per month versus lifespan (weeks). (B) Rate of mean telomere length shortening (kb/month) versus the rate of increase in the percentage of short telomeres (per month). (C) Rate of mean telomere length shortening (kb/month) versus the rate of increase in the percentage of short telomeres (<15 kb) per month. The slope of each regression line is indicated. See also Figures S1, S2, S3, S4, and S5. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure S1 Telomere Length at Different Ages and its Correlation with Longevity, Related to Figure 2 (A) The mean telomere length was determined by HT-QFISH on white blood cells from WT mice at the indicated time points. Linear regression lines were calculated for the values obtained (kb) versus lifespan (weeks). The slope of each regression line is indicated. (B) The percentage of short telomeres (<15 kb) was determined by HT-QFISH on white blood cells from WT mice at the indicated time points. Linear regression lines were calculated for the values obtained (%) versus lifespan (weeks). The slope of each regression line is indicated. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure S2 Rate of Increase in the Percentage of Short Telomeres and Rate of Mean Telomere Shortening at Different Age Intervals, Related to Figure 2 (A) The rate of increase in percentage of short telomeres (<15 kb) was determined by HT-QFISH on white blood cells from WT mice at the indicated time points. Values are given as average ± SEM. From left to right (average ± SEM): 0.8719 ± 0.1702, 0.6589 ± 0.3524, 0.5787 ± 0.3407, 0.9236 ± 0.2353, 1.336 ± 0.5503, 0.06455 ± 0.7752. (B) The rate of mean telomere length was determined by HT-QFISH on white blood cells from WT mice at the indicated time points. Values are given as average ± SEM. From left to right (average ± SEM): 1.65 ± 0.4148, 0.74 ± 0.368, 0.4256 ± 0.1863, 1.395 ± 0.4224, 1.71 ± 0.6954, 0.1043 ± 0.6611. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure S3 The Rate of Increase in the Percentage of Short Telomeres and the Rate of Telomere Shortening Is a Predictor of Mouse Longevity, Related to Figure 2 (A and B) The rate of increase in percentage of short telomeres (<15 kb; A) and mean telomere length (B) were determined by HT-QFISH on white blood cells from the indicated mice cohorts at the indicated time points (see Figure 1). Linear regression lines were calculated for the values obtained for the rate of increase in the percentage of short telomeres and for the rate of mean telomere length shortening (kb/month) versus lifespan (weeks). (A) Rate of increase in the percentage of short telomeres (<15 kb) per month versus lifespan (weeks). (B) Rate of mean telomere length shortening (kb/month) versus increase in the percentage of short telomeres (per month). Only results from TgTERT mice for which the four time points were measured are plotted. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure S4 Correlation between Rates of Telomere Shortening and Weight, Related to Figure 2 (A) Linear regression lines were calculated for the net weight gain of each individual mouse (grams) versus lifespan (weeks). The slope of each regression line is indicated. (B) Linear regression lines were calculated for the net weight gain of each individual mouse (grams) versus the rate of increase in the percentage of short telomere (%/month). The slope of each regression line is indicated. (C) Linear regression lines were calculated for the net weight gain of each individual mouse (grams) versus the rate of mean telomere shortening (kb/month). The slope of each regression line is indicated. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Figure S5 Mice with Faster Rates of Increase in Short Telomeres Present the Same Pathologies, but at an Earlier Onset than Those with Lower Rates of Increase in Short Telomeres, Related to Figure 2 (A) Percentage of WT mice with the indicated tumors or degenerative pathologies at their time of death, belonging to the lower (<0.24%/month) or higher (>0.92%/month) quartiles of the increase in the percentage of short telomeres. The mean lifespan of the different quartiles is also represented. (B) Percentage of TgTERT mice with the indicated tumors or degenerative pathologies at their time of death, belonging to the lower (<0.012%/month) or higher (>0.68%/month) quartiles of the rate of increase in the percentage of short telomeres. The mean lifespan of the different quartiles is also represented. Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions

Cell Reports 2012 2, 732-737DOI: (10.1016/j.celrep.2012.08.023) Copyright © 2012 The Authors Terms and Conditions