Fig 1.
(A) Immunohistochemical staining of FoxO1 protein in the mouse uterus throughout days 1–8 of pregnancy. D5 and D6 demonstrate the implantation sites where D5-i and D6-i show inter-implantation sites on days 5 and 6 of pregnancy. Rectangles indicate higher magnifications in the lower row images. Negative control staining indicates the specifity of FoxO1 immunohistochemistry. (B) Western blot and relative densitometric analysis of FoxO1 during peri-implantation period in mice. Mouse brain tissue was used as positive control for FoxO1 expression. GAPDH levels are shown as a loading control. Experiments were repeated at least three times. LE: Luminal epithelium, GE: Glandular epithelium, E: Endothelium, B: Blastocyst, EM: Embryo, PDZ: Primary decidual zone, SDZ: Secondary decidual zone, CE: Chorionic ectoderm, AM: Amnioblasts, EP: Epiblast. NC: Negative control. Scale bars: 50 μm.
Fig 2.
(A) Immunolocalization and H-score analyses of FoxO1 at different time intervals through days 4 and 5 of pregnancy [WOI in mice; Day 4 (09:00 AM), Day 4 (19:00 PM), Day 5 (09:00 AM) and Day 5 (19:00 PM)]. (B) Immunohistochemical expression of FoxO1 on day 5 (09:00 AM) and (19:00 PM) of pregnancy at implantation site and inter-implantation site on day 5 (19:00 PM). Note that, specifically at the region of blastocyst attachment, FoxO1 expression was cytoplasmic in luminal epithelium (dashed rectangle). LE: Luminal epithelium; GE: Glandular epithelium; S: Stroma; E: Endothelium; B: Blastocyst; PDZ: Primary decidual zone; L: Lumen. Scale bars: 50 μm.
Fig 3.
(A) Immunohistochemical expression of FoxO1 in the mouse uterus during pseudopregnancy on days 1, 2, 3, 4, 5, 6 and 8 (PP-D1, PP-D2, PP-D3, PP-D4, PP-D5, PP-D6, and PP-D8). Rectangles indicate higher magnifications in the lower row images. (B) H-score analyses of FoxO1 during pseudopregnancy uterus in mice. LE: Luminal epithelium, GE: Glandular epithelium, E: Endothelium. Scale bars: 50 μm.
Fig 4.
(A) FoxO1 expression in the mouse uterus under artificial decidualization on days 5, 6 and 8. ADC-D5, ADC-D6, and ADC-D8, uninjected uterine horn for control; AD-D5, AD-D6, and AD-D8, uterine horn under artificial decidualisation. Rectangles indicate higher magnifications in the lower row images. (B) A representative photo showing the deciduoma on day 5, 6 and 8 pseudopregnant mice under artificial decidualization treated with oil. (C) H-score analyses of FoxO1 in the mouse uterus under artificial decidualization. LE: Luminal epithelium, GE: Glandular epithelium, E: Endothelium. Scale bars: 50 μm.
Fig 5.
FoxO1 expression in ovariectomized, delayed implanting uteri before and after E2 activation.
(A) Delayed implantation (Delayed) and activation of delayed implantation by estrogen (Activated). (B) H-score analyses of FoxO1 in the mouse uterus under delayed implantation and activation. LE: Luminal epithelium, GE: Glandular epithelium, E: Endothelium. Scale bars: 50 μm.