Hundred milligrams of the quadriceps muscle (wet) were separated and homogenized with PBS containing antiproteases (0.1 mM PMSF, 0.1 nM hydrochloric benzethonium, 10 mM EDTA and 20 Ki aprotinin A) and 0.05% Tween 20. The samples were centrifuged for 10 min, at 10,000 rpm and at 4°C. The supernatant was used for the ELISA assay with a 1: 4 dilution. The ELISA assay was performed according to the manufacturer’s instructions (R&D System) and quantified from the 492 nm wavelength acquired in a plate reader (Spectramax plus 384, Molecular Devices, United States).

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