Fig. 4

Inhibition of mTOR signaling by Rapalink-1 at the 8-cell stage impaired the morula to blastocyst transformation and decreased the differentiation of trophectoderm lineage. A Schematic of experimental setup whereby 8-cell embryos were treated in mTOR inhibitor. B The percentage of morula and blastocyst after 24 h cultured in the different treatment conditions. C Representative immunofluorescence images of CDX2 and SOX2 in blastocyst from the RPL treatment and Con group. Scale bars, 20 μm. D A significantly decreased the total cell number in mTOR inhibitor treated embryos (n = 7). E Rapalink-1 treatment decreased the ratio of TE cells. F Representative immunofluorescence images of SOX17, NANOG and CDX2 expression in blastocyst derived from RPL and Con group (n = 7). G The cell number of total, SOX17⁺, NANOG⁺ and CDX2⁺ of blastocysts in the RPL and Con group (n = 7). H Percentage of SOX17⁺, NANOG⁺ and CDX2⁺ cells of the blastocyst in Con and RPLgroup (n = 7). RPL: Rapalink-1 and Con: Control. Error bars are mean ± SEM. ^*p < 0.05, ^** p < 0.01, n.s., not significant (p > 0.05)