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Total RNA was then used to synthesize complementary DNA, followed by the performance of real-time PCR using an SYBR Green I technology and MxPro-Mx3000P QPCR machine

For experiments, cells were cultured on round coverslips in 12-well plates and then stabilized for 30 min with Dulbecco's modified Eagle's medium supplemented with 5 mg/mL apotransferrin, 1 mg/mL bovine serum albumin, 25 mM HEPES (pH 7.4), 2 mM glutamine, 100 units/mL penicillin, and 100 mg/mL streptomycin (serum-free medium)

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Thats in part because you can have high total testosterone but low free testosterone, which is the type your body actually uses

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