1. Assisted reproductive technologies do not increase risk of abnormal methylation of PEG1/MEST in human early pregnancy loss 
Hai-Yan Zheng, M.D., Xiao-Yun Shi, M.D., M.Sc., Fang-Rong Wu, M.D., Ya-Qin Wu, M.D., Le-Le Wang, M.D., Shi-Ling Chen, M.D., M.Sc. 
Fertility and Sterility 
Volume 96, Issue 1, Pages e2 (July 2011)
DOI: /j.fertnstert
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
(A–C) Pyrosequencing results of PEG1 in controls and chorionic villus. “T%” of two sites marked with an asterisk (∗) represents the bisulfite conversion efficiency. (A) Unmethylated pattern in sperm DNA (0.85% methylation). (B) Methylated pattern in oocyte DNA (94.8% methylation). (C) Normal methylation profile in villus (53.5% methylation). (D) Box plots show the distribution of methylation values for the four experimental groups. Median values are shown as horizontal lines. Bottom of the box indicates the 25th percentile, top the 75th percentile. Outliers are shown as asterisks (cases 1, 41, and 129). (E) ROC curve analysis demonstrates a positive correlation between methylation percentage of PEG1 and early spontaneous abortion rate. Area under the curve was (95% confidence interval, 0.592–0.764).
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Bisulfite-PCR sequencing for cases 1, 41, 57, 129, and 136. Vertical bars signify CpG sites. The region between two inner arrows, including four CpG sites (∗), represents the sequences assessed by pyrosequencing. Each row represents a unique methylation profile within the pool of clones sequenced, with the frequency of that methylation state shown to the right of each row. Closed and open circles represent methylated and unmethylated CpGs, respectively. Results are summarized in Table 2.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

4. Supplemental Figure 1
Location of PEG1 gene's sequencing primer. There are four methylated CpG sites and two conversion efficiency sites. “T” represents methylated CpG site, and “C” represents conversion efficiency site.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

5. Supplemental Figure 2
Representative example of MS-PCR analysis of PEG1 in chorionic villus from four experimental groups. Paternal-specific MS-PCR products (300 bp) are shown on the left, maternal-specific products (300 bp) on the right. Lane 1, spontaneous abortion after ART. Lane 2, multifetal reduction after ART. Lane 3, spontaneous abortion of natural pregnancy. Lane 4, induced abortion of natural pregnancy.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions