A vaginal smear was performed at approximately 9:00 am. Briefly, 10 µL of saline was pushed into the vagina and drawn with a micropipette 3 times. Then, saline containing exfoliated vaginal cells was smeared on a slide. After air drying, the smear was fixed with 75% ethanol. HE staining was performed to evaluate the cell types. The oestrous cycle was read and recorded for each mouse.
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