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At 30 min (for pERK1/2 staining) or 90 min (for c-Fos staining) after injection, mice were transcardially perfused with phosphate buffer including 4% paraformaldehyde and 0.2% picric acid under anesthesia

doi: 10.1371/journal.pone.0296733 287

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This process contributes to estrogen excess and blocks the effects of true estrogen

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