DEV023747 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. (A) Methylation of H3-K9 in enucleated GV oocytes. Somatic chromosomes with histone demethylation can commence meiotic division and reach an MII-like stage. Me-H3-K9 is shown in green; microtubules are stained for β-tubulin in red. (B) Expression of histone H3 phosphorylation at serine 28 (P-H3-S28) in somatic and oocyte chromosomes in intact GV oocytes injected with somatic nuclei. P-H3-S28 is shown in green. (C) Intact GV oocyte injected with somatic cell nuclei and cultured for 17 hours. The somatic cell chromosomes showed a different morphology from those of oocyte. The oocyte chromosomes were at MII but the somatic chromosomes resembled those of fully grown GV oocytes and arrested at this stage. Scale bar: 20 µm (OC, oocyte chromosome; SC, somatic chromosome).

• Supplemental Figure S2 -

  Fig. S2. Intact cumulus cell nuclei (A) and cytoplasmic lysate-treated cumulus cell nuclei (B) were injected into intact MII oocytes and cultured without activation for 2 hours. The demethylation of H3-K9 in the lysate-treated cumulus group was maintained, as seen in somatic chromosomes (B′). Then, oocytes were activated and cultured for 6 hours to compare histone H3 methylation in the oocyte and in cumulus-injected pronuclei (C′ and D′). Methylation of H3-K9 is shown in green and nuclear membranes are stained by lamin B in red (OC, oocyte chromosomes; SC, somatic chromosomes; OP, oocyte pronucleus; SP, somatic pronucleus). Scale bar: 20 µm.

• Supplemental Figure S3 -

  Fig. S3. Levels of Oct-4 and Cdx2 protein expression in expanded blastocysts derived from normally fertilized embryos, embryos produced by nuclear transfer with intact cumulus cells and embryos produced by nuclear transfer with cumulus cells treated with GV oocyte cytoplasmic lysates. Blastocysts were immunostained for Oct4 and Cdx2 proteins.