Fig. 2: The temporal expression and function of ATF3 in the endometrium.

A, B Immunohistochemistry analysis with an anti-ATF3 antibody. Proliferative and mid-secretory phase endometrial tissue samples from normal fertile women. The negative control (NC) was nonspecific rabbit serum. The H-score of ATF3 expression in the endometrial stromal cells was calculated with IHC-Profiler and ImageJ (Scale bar = 50 µm, n = 3, *P < 0.05). C Expression of ATF3 in proliferative and early-, mid-, and late-luteal phase endometrium. Each bar represents an individual biopsy. The data were retrieved from microarray data deposited in the Gene Expression Omnibus (GDS2052). D, E ATF3 expression pattern analyzed with a recent single-cell RNA-seq analysis in stromal cells (D) and all cells (E). The data were retrieved from microarray data deposited in the Gene Expression Omnibus (GSE111976). F The expression pattern of ATF3 in hESCs treated with 0.5 mM 8-Br-cAMP and 1 μM MPA (M + A) for different periods of time (0, 0.5, 1, 2, 4, 8, or 16 h) was evaluated by qPCR. *P < 0.05, **P < 0.01. G The expression pattern of ATF3 in hESCs treated with 0.5 mM 8-Br-cAMP and 1 μM MPA (M + A) for different periods of time (0, 1, 3, 6, 12, 24, 48, or 72 h) was evaluated by western blot. H GSEA results showed that the ATF3-regulated gene term was enriched during in vitro decidualization. I, J hESCs were transfected with siATF3 or siCtl for 48 h and then treated with a decidualization stimulus. The expression and secretion of PRL were measured by qPCR and ELISA, respectively. *P < 0.05, **P < 0.01 compared with the CTL/M + A group (8-Br-cAMP+MPA). K Immunofluorescence was performed to analyze the morphological transformation of hESCs.