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The following day, DPX mounting media for histology (Sigma-Aldrich, Castle Hill, NSW, Australia) was dropped onto each section prior to mounting with coverslips (KNITTEL Coverglass, ProSciTech, Kirwan, Queensland, Australia)

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the incubates were then spun through Bio-Gel P6 spin columns (400 packed volume) for 1 min to separate free and bound 3 H-Hinge and the eluent was collected