After sacrifice, mouse muscle tissue was washed with 0.9% saline and fixed in 10% formalin. Tissues were embedded in paraffin and cut into 4 μm thick slices for morphological and pathological evaluations. Immunohistochemical (IHC) staining of tissues involved the use of the Leica antibody to myosin heavy chain fast (WB-MHCf) and myosin heavy chain slow (WB-MHCs). By using automated BondMax with double staining, WB-MHCf and WB-MHCs epitope retrieval involved the use of ER2 (AR9640) retrieval solution for 30 min once, followed by incubation with WB-MHCf and WB-MHCs antibodies with diluent 100X for 30 min. The detection kit used was the Bond Polymer Refine Detection (DS9800) (incubation with post-primary for 8 min, polymer for 8 min, and 3′3′-diaminobenzidine for 5 min) and Bond Polymer Refine Red Detection (DS9390) (incubation with post-primary for 20 min, polymer for 30 min, red for 10 min, and haematoxylin for 5 min). Finally, results were examined under a light microscope equipped with a CCD camera (BX-51, Olympus, Tokyo) by a veterinary pathologist.

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