Research Paper Volume 15, Issue 5 pp 1257—1278

Figure 2. Validation of sperm ageDMRs by bisulfite pyrosequencing. Scatter plots showing the correlations between average regional methylation (y-axis in %), determined by bisulfite pyrosequencing, and donor age (x-axis in years) in 94 human sperm samples. The blue dots represent pyrosequencing measurements for an ageDMR (identified by RRBS) in EEF1A2, the red dots for an ageDMR in MBD3, the green dots for an ageDMR in PRAM1, and the yellow dots for an ageDMR in PRKAR2A. Consistent with the results of RRBS, the regression lines of all analyzed regions indicate a significant (also see Supplementary Table 3) loss of methylation with age. The correlations remain virtually unchanged when excluding the 72-year-old sample from analysis.