Fig. 3

Assessment of the cell fate specification in blastocyst stage and the quality of blastocysts derive from mTOR inhibition embryos. A Representative 3D images of CDX2 and SOX2 immunofluorescence of blastocysts developed from the mTOR inhibitor treatment and without treated conditions. SOX2 positive cells represent the ICM (red), while CDX2 positive cells represent the TE (green). Nuclei were stained with Hoechst33342 (blue). Scale bars, 20 μm. B The number of TC, ICM, and TE of blastocysts in the RPL and Con group (n = 7). C The percentage of ICM cells and TE cells in the RPL and Control group (n = 7). D Representative immunofluorescence images of SOX17, NANOG and CDX2 of blastocyst derivation from the RPL treatment and Con group. Scale bars, 50 μm. E The cell number of total, SOX17⁺, NANOG⁺ and CDX2⁺ of blastocysts in the RPL and Con group (n = 7). F The percentage of SOX17⁺, NANOG⁺ and CDX2⁺ cells in each group (n = 7). G Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining of the blastocysts derived from the RPL treatment and Control group. Scale bars, 50 μm. H The graph showing the quantification of the apoptosis cells (n = 10). RPL Rapalink-1, Con Control. TC total cell number, ICM inner cell mass, TE trophectoderm. Error bars are mean ± SEM. ^*p < 0.05, ^** p < 0.01, n.s., not significant (p > 0.05)