Fig. 5: Abnormal activation of the WNT signaling pathway impaired decidualization both in vitro and in vivo.

a Measurement of luciferase activity in TOPflash-transfected and FOPflash-transfected hESCs treated with LiCl or control during decidualization (n = 3). b IF assay of β-catenin expression in hESCs treated with LiCl or control (n = 3); scale bar = 50 μm. c qRT-PCR analysis of PRL and IGFBP1 expression in the control and LiCl-treated groups treated with E2, MPA, and 8Br-cAMP for 4 days (n = 3). d Schematic representation of the natural decidualization procedures with LiCl treatment. e Gross morphology of D8 uterine from the PBS and LiCl treatment (60 mg/kg) groups; scale bar = 1 cm. f Hematoxylin-eosin staining of D8 uterine from the PBS and LiCl treatment (60 mg/kg) groups; scale bar = 1 mm. g Number of implantation sites from the PBS and LiCl treatment (60 mg/kg) groups (n = 5). h Weight of implantation sites from the PBS and LiCl treatment (60 mg/kg) groups (n = 5). i Immunostaining of Ki67 expression in D8 implantation sites from the PBS and LiCl treatment (60 mg/kg) groups, and the ratio of the length of Ki67-positive site to the Ki67-negative site (n = 3); scale bar = 200 μm. j qRT-PCR analysis of Prl8a2 expression in D8 uterine from the PBS and LiCl treatment (60 mg/kg) groups (n = 5). k Schematic representation of the stimulated decidualization procedures with LiCl treatment. l Gross morphology of the unstimulated or stimulated uterine from the PBS and LiCl treatment (60 mg/kg) groups; scale bar = 1 cm. m Hematoxylin-eosin staining of the unstimulated or stimulated uterine from the PBS and LiCl treatment (60 mg/kg) groups; scale bar = 1 mm. n Ratio of stimulated to unstimulated uterine weight from the PBS and LiCl treatment (60 mg/kg) groups (n = 5). o qRT-PCR analysis of Prl8a2 expression in stimulated uterine from the PBS and LiCl treatment (60 mg/kg) groups (n = 5). Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Student’s t test and one-way ANOVA.