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Ovaries were removed from mice and punctured with a 25-gauge needle to isolate granulosa cells. Granulosa cell/oocyte mixture was obtained by passing the tissue lysate through a 70 μm cell strainer (BD Falcon, USA). To further purify granulosa cells, the granulosa cell/oocyte mixture was then filtered through a 40 μm cell strainer (BD Falcon, MA, USA), which will separate granulosa cells from oocytes. The purity of granulosa cells was determined by staining isolated cells with an anti-FSHR antibody (Thermo Fisher CL594-22665).
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