For analysis of cells embedded in soft agar, cells (10 5 ) were suspended in 1 mL of 0.33% noble agar, seeded in the bottom of 15-mL polypropylene tubes, cooled for 5 min on ice, and then kept at 37 C in a humidified incubator in the presence of 5% CO 2 for 24 h72 h
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At the end of the treatment period, after euthanizing the animals, serum levels of LH, FSH, testosterone, insulin and glucose were analyzed, and histological, immuno-histochemical (IHC), and molecular examinations of the ovaries were performed
