Fig 4. Ectopic lesions are decreased in PUGKO mice compared to controls.

A. Creation of tdT PUGKO mouse model with daily injections of progesterone from postnatal day 2–10 to knock down secretome factors was based on mouse model that results in infertility in female mice. P4 = progesterone B. Oviduct tissue and endometrium from progesterone treated (PUGKO) and vehicle control donor mice were collected and processed, then injected into WT C57Bl/6J recipient mice. Due to the fluorescent nature of the tdT mouse tissue, all tissue that implanted in the recipient mice could be identified using a red filter with a green light exposure. C -D. Immunohistochemistry (IHC) staining for FOXA2 was performed to assess the response of pups to progesterone treatment (PG TX). This was measured by comparing the counted IHC FOXA2 stained uterine glands of microscopic images in 4 high power fields (HPF) of 40 magnification in PUGKO tdT (D) vs WT (C) Scale Bar = 100 μm. PG treatment in PUGKO mice decreases the expression of FOXA2 almost 33 times compared to the non-PG TX mice. E. Lesion count in mice with endometrial tissue from tdT mice ± progesterone on postnatal day 2–10 (PUGKO). There are significantly less lesions when mice recieve progesterone as pups (* = p<0.05). F. Lesion development using PUGKO endometrium and secretome factors (provided by synchronized WT endometrium). PUGKO reduces implantation factors and associates with fewer endometrial lesion growths. WT endometrium appears to restore implantation of PUGKO endometrium. There is no statistically significant difference when PUGKO endometrium is co-injected with WT endometrium. G. PUGKO does not significantly alter the number of oviductal lesions. The progesterone induced blocking of uterine glands in PUGKO mice did not statistically affect the number of oviductal lesions compared to controls in vehicle mice, since oviduct does not produce secretome factors. (Student t-test.) E: Estrogen, P: Progesterone.