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. 2015 Apr;185(4):1104–1113. doi: 10.1016/j.ajpath.2014.12.019

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Immunohistochemical (IHC) analysis of downstream signaling molecules. A and B: Positive IHC staining with p-Gsk3β (A) and p-Pras40 (B) antibodies in uteri with complex atypical hyperplasia (CAH; arrow) as compared to normal epithelium (arrowheads). C: Loss of nuclear FoxO1 staining in CAH (arrows) compared to the intense nuclear staining of normal epithelium (arrowheads). D: Loss of nuclear FoxO1 in carcinoma. E: Immunoblot analysis depicting subcellular localization of FOXO1 in endometrial cancer cell lines. Lamin A/C and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as controls to determine relative purity of nuclear (N) and cytoplasmic (C) extracts. WT, wild type.