Immediately after culling, blood was collected from mice by cardiac puncture with a syringe and needle pre-treated with heparin. Plasma was separated by centrifugation and stored at −80 °C. Testosterone was measured using an ELISA kit (DEV9911; Demeditec Diagnostics, Kiel, Germany). LH was measured using an in-house designed ELISA as previously described54 (link). Samples for each treatment group or age group were analysed together.
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