Changes in Histone H3 Lysine 36 Methylation in Porcine Oocytes and Preimplantation Embryos
Figure 7
Changes in H3K36me3 status in porcine IVF and SCNT embryos.
(A) Sperm, MII stage oocytes coincubated with sperm in mTBM medium for 1 h (IVF 1 h), and IVF embryos cultured in PZM-3 medium for 18 h (IVF) were immunostained with the anti-H3K36me3 antibody. (B) A single fetal fibroblast cell injected into the perivitelline space of a denuded oocyte without activation (Embedded), a nuclear-transferred embryo cultured in PZM-3 for 1 h post activation (1h), a nuclear-transferred embryo cultured in PZM-3 for 18 h after activation (18h), and other stages of SCNT embryos were immunostained with the anti-H3K36me3 antibody. The antibody was localized with a FITC-conjugated secondary antibody (green). DNA was stained with DAPI (blue). The arrowhead indicates sperm and the arrow indicates the nuclei of donor cell. Scale bar = 20 µm. (C) Relative intensities of fluorescence signals from H3K36me3. Total 41 embryos were analyzed in triplicate in this experiment. Bars represent least-squares showed the standard error in each group. P-values <0.05 were considered statistically significant. a,b,c Values represent least-squares with different superscripts are significantly different (P<0.05).