Copy number analysis was performed following the protocol reported previously with minor modifications 91. Briefly, MCF7 cells cultured in stripping medium for three days were treated with estrogen (10-7 M) for 6 h, and 1.5 × 106 cells were then used for copy number analysis. Absolute quantification was performed using 2-fold serial dilutions of the reference standard, and Ct value from qPCR analysis versus the dilution factor was plotted, fitting the data to a straight line. The standard curve was used for extrapolating the number of molecules of interest.

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