1. Volume 1, Issue 5, Pages 506-515 (May 2012)
Rapid Mitochondrial DNA Segregation in Primate Preimplantation Embryos Precedes Somatic and Germline Bottleneck 
Hyo-Sang Lee, Hong Ma, Rita Cervera Juanes, Masahito Tachibana, Michelle Sparman, Joy Woodward, Cathy Ramsey, Jing Xu, Eun- Ju Kang, Paula Amato, Georg Mair, Ralf Steinborn, Shoukhrat Mitalipov 
Cell Reports 
Volume 1, Issue 5, Pages (May 2012)
DOI: /j.celrep
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2. Cell Reports 2012 1, 506-515DOI: (10.1016/j.celrep.2012.03.011)
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3. Figure 1
Schematic Model Demonstrating mtDNA Segregation and Bottleneck in Primates
Heteroplasmic rhesus monkey oocytes with equal mixture of two wild-type mtDNA haplotypes were constructed and mtDNA transmission to preimplantation embryos, fetuses, and germ cells was followed. We demonstrate rapid segregation of mtDNA variants between daughter blastomeres in preimplantation embryos. However, fetuses and ESCs derived from these embryos shifted toward homoplasmic conditions. This bottleneck suggests that possibly a few cells within an ICM contribute to the somatic cell lineage of embryo proper. In contrast, individual fetal oocytes (F2) showed a wide range of heteroplasmy. This model also implies that the majority of ICM cells may contribute to the germline.
See also Table S4.
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4. Figure 2
Segregation of mtDNA in Individual Blastomeres of the 2-, 4-, and 8-Cell Embryos
(A–E) Segregation of alien mtDNA between daughter blastomeres of individual (A) 2-cell, (B) 4-cell, and (C) 8-cell embryos expressed by range (maximum or minimum alien mtDNA values minus the mean median values). Comparison of mtDNA dispersal between 2-, 4-, and 8-cell embryos based on the (D) coefficient of variation and (E) range. Different letters indicates p values <0.05.
(F) MtDNA segregation between the ICM and TE in individual blastocysts. The mean alien mtDNA in ICM and TE were 56.8% ± 10.7% and 58.4% ± 9.0%, respectively. Data are represented as mean ± SD.
See also Tables S1–S3.
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5. Figure S1
Restriction Fragment Length Polymorphism Analysis of mtDNA in Organs and Tissues of Heteroplasmic Fetuses and ESCs, Related to Table 2
(A) MtDNA heteroplasmy detected by restriction fragment length polymorphism (RFLP) in tissues from the female fetus. Resident mtDNA digested with KpnI and alien mtDNA treated with NheI produced fragments of 319 bp + 224 bp and 417 bp + 126 bp length, respectively. RFLP analysis showed that the major mitochondrial haplotype in all tissues from this fetus was alien while the resident mtDNA was not detected except in liver and heart.
(B) Heteroplasmy in tissues from the male fetus. RFLP assay with unique enzymes (HindIII for resident mtDNA and EcoRV for alien mtDNA) detected presence of both variants in this fetus. However, alien mtDNA bands were noticeably weaker.
(C) Heteroplasmy in ESC lines produced from heteroplasmic embryos. Two cell lines (Hetero-1 and −3) carried alien mtDNA while Hetero-2 contained resident haplotype. For quantifications, both alien and resident mtDNA were mixed at various ratios (0, 10, 30, 50, 70, 90, and 100%).
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6. Figure S2 Gender Determination in ST Blastocysts by TE Biopsy and PCR
DNA was extracted from individually biopsied blastocysts and amplified by PCR. Male DNA samples produced two bands (1149 bp and 771 bp) and female DNA produced only one band (1149 bp). Blood DNA from rhesus monkey males (M) and females (F) was used as controls. For negative control (N) no DNA template was used. Samples in lanes 1, 2, 3, 5 and 7 were females and lanes 4 and 6 were indicated male blastocysts.
Cell Reports 2012 1, DOI: ( /j.celrep )
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