Preparation of tumor lysate and western blotting

Western blot samples were prepared as described²⁵. Briefly, tissue samples stored at −80 °C were thawed in RIPA buffer (200 μL to 400 μL) supplemented with proteinase- and phosphatase-inhibitor cocktail (Sigma), then homogenized with an electrical homogenizer followed by short sonication to form a homogeneous tissue lysate. The lysate solution was centrifuged at 18,000 × g for 30 min at 4 °C. Protein concentration was measured with the BCA assay (Thermo Fisher). Cell lysate of fallopian tube cells FT237 and FT246 were obtained from Dr. Analisa Difeo (University of Michigan, MI). Thirty to forty μg protein per sample was subjected to SDS-PAGE analysis. The lysates of ovarian surface epithelial cells and fallopian tube cell 15–20 µg protein was used for WB analysis. Proteins were then electro-transferred to methanol activated immobilon-FL PVDF membranes (EMD Millipore). Membranes were blocked with Starting Block (Thermo Fisher) for 1 hr at room temperature and incubated with primary antibodies overnight at 4 °C. Dylight 800-conjugated secondary antibodies were used for detection (Thermo Fisher, 1:5000, 5% milk, 1 hour, RT) of fluorescent signals in Odyssey Imaging Systems (LI-COR Biosciences).