Fig. 5

Transcription profiles of blastocysts and key gene terms enriched in mTOR inhibitor treatment during 8-cell to blastocyst development. A Volcano plot displaying the DEGs in RPL blastocysts. B Bar chart illustrating the enriched GO terms associated with the significantly downregulated transcripts in RPL blastocysts identified by RNA-Seq. C Bar chart illustrating the enriched GO terms associated with the significantly upregulated transcripts in RPL blastocysts. D GSEA plots showing trophectodermal cell differentiation enriched in RPL-blastocyst (left) and Con-blastocyst (right). NES, normalized enrichment score; FDR, false discovery rate. E Heatmap illustrating the relative expression level (row-scaling) of the genes associated with trophectodermal cell differentiation between RPL and Control blastocyst. F Bar chart illustrating the top ten enriched KEGG terms or canonical pathways in RPL blastocyst. G GSEA profiles showing the most significant enrichment of gene sets associated with lysosomes in RPL blastocysts (RPL vs Con). H The heatmap shows the expression pattern of lysosome-related genes in RPL and Con blastocysts. I Representative immunofluorescence images of lamp1 (lysosomal associated membrane protein 1) and LC3β (microtubule-associated protein 1 light chain 3β) in RPL and Con embryos. Bar, 50 μm. J The normalized intensity of lamp1 between Con and RPL group embryos (n = 10). K The normalized intensity of LC3β between Con and RPL group embryos (n = 10)