Extended Data Fig. 1: The mid-secretory endometrium exhibits different phenotypes and gene expression between young and middle-aged patients.

a, The percentage of Ki67-positive cells in endometrial luminal epithelial and stromal cells in the mid-secretory phase (n = 3). b, Relative protein levels of IGFBP1 in human endometrial stromal cells during induced decidualization (n = 3). c,d, Histochemical scoring assessment (H-score) of PGR and ERα in the mid-secretory endometrium (n = 3). e, Relative protein levels of PGR and ERα in the mid-secretory endometrium (n = 5). f,g, Protein levels of PGR and ERα in human mid-secretory endometrial stromal cells (n = 4). h, Relative mRNA levels of PGR and ESR1 in human mid-secretory endometrial stromal cells (n = 4). i,j, Protein levels of PGR and ERα in human mid-secretory endometrial epithelial cells (n = 4). k, Relative mRNA levels of PGR and ESR1 in human mid-secretory endometrial epithelial cells (n = 4). l, PCA plot of human mid-secretory endometrial RNA-seq data. m, Volcano plot illustrating gene expression changes between the young and aging mid-secretory endometrium (|log2FC| > 1, P.adj < 0.05). n, Heatmap showing gene expression of differentially expressed genes (DEGs) between the young and aging mid-secretory endometrium. o, FPKM of LIF, SOX4, FGF1, WNT2, IL7, IHH in the human mid-secretory endometrium (n = 10 and n = 8 for the young and middle-aged groups, respectively). The adjusted P value was determined by DESeq2. Whiskers represent upper quartile + 1.5 interquartile range (IQR) and lower quartile − 1.5 IQR. p, Pathway enrichment analysis of upregulated DEGs in the aging mid-secretory endometrium. Statistical analysis was performed by two-sided unpaired Student’s t-test or Mann–Whitney U rank-sum test. Data are presented as mean ± s.d. All replicates were biological replicates. FC, fold change; mid-aged, middle-aged; ns, not significant; P.adj, adjusted P value; PC, principal component; PCA, principal component analysis.

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