Research Paper Volume 12, Issue 3 pp 2680—2697

Figure 3. Methionine does not shorten the long lifespan achieved by calorie restriction. (A) The long lifespan from feeding diluted bacteria is not significantly affected by 5mM methionine. Ad libido (AL) and CR conditions were achieved by plating 1X10¹¹/mL and 1X10⁸/mL OP-50 bacteria, respectively, on solid nematode growth (NG) medium containing carbenicillin and kanamycin. Worms were raised on NG medium supplemented with or without 5mM methionine (Met) throughout life with frequent transfer to new plates to avoid contamination by progenies. Number of deaths and lives were recorded every other day. Data from 3 independent experiment were pooled and plotted by using Graphpad Prism software. P values were obtained by log-rank test: CR (met) vs. CR, not significant; CR(met) vs. AL, P<0.0001. See Supplementary Table 1 in Supplemental Information (SI) for more information. (B) The long lifespan from CR model (eat-2 mutant) is not significantly affected by 5mM methionine. N2 wild-type and eat-2(ad1116) worms were raised on normal NG medium with sufficient OP-50 bacterial food and also supplemented with and without 5mM methionine (Met). Number of deaths and lives were recorded every other day and with frequent transfer to new plates to avoid contamination by progenies. Data from 3 independent experiment were pooled and plotted by using Graphpad Prism software. P values were obtained by log-rank test: eat-2(met) vs. eat-2, not significant; eat-2(met) vs. N2, P<0.0001. See Supplementary Table 2 in Supplementary Information (SI) for more information.