Fig. 3.

Effect of Notch signaling inhibition during IVM on early porcine embryogenesis. A Representative images of blastocysts developed from control and RO4929097-treated groups after parthenogenetic activation. Bar = 200 µm. B Cleavage and (C) blastocyst formation rates of embryos from control and RO4929097-treated groups after parthenogenetic activation (0: n = 219; 5: n = 161; 10: n = 138; 20: n = 113). D Representative images of blastocysts at four different stages: early blastocyst (EB), middle blastocyst (MB), large blastocyst (LB), and expanded blastocyst (ExB). Bar = 50 µm. E Proportions of various blastocyst stages in the control and RO4929097-treated groups (0: n = 133; 5: n = 87; 10: n = 51; 20: n = 40). F Representative nuclear-stained images of blastocysts in the control and RO4929097-treated groups. Bar = 50 µm. G Quantification of total cell numbers in the control and RO4929097-treated groups (n = 20 per group). H Representative images of TdT-mediated dUTP nick-end labeling (TUNEL)-stained blastocysts. Bar = 50 µm. Embryos were subjected to TUNEL (green, white arrow) and nuclear staining (blue). I Quantification of the proportion of apoptotic cells in the indicated groups (n = 40 per group). J qRT-PCR-assayed transcript levels of apoptosis-related genes (BAX and BCL2L1) in blastocysts in the indicated groups (n = 3 per group). K Representative images of CDX2 staining in blastocysts. Bar = 50 µm. L Quantification of cell numbers in the inner cell mass and trophectoderm in the indicated groups (n = 33 per group). M qRT-PCR-assayed transcript levels of developmental potential-related genes (POU5F1, NANOG, SOX2, CDX2, TEAD4, and GATA3) in blastocysts in the indicated groups (n = 3 per group). Data are derived from at least three independent experiments, and different superscripts indicate significant differences (P < 0.05). Con, control; RO, 10 µM RO4929097