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Cells were treatment with protein synthesis inhibitor Cycloheximide (MCE, #HY-12320, 10 μg/ml) alone or combined with proteasome inhibitor MG132 (MCE, #HY-13259, 20 μM) for 0, 2, 4, or 6 h. Cell lysates were collected and the protein levels of ERα were analyzed by WB. The ERα density was quantified using Quantity One Software (v4.62, Bio-Rad) and used to calculate the half-life time (t1/2) of the ERα proteins.
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