Muscle Fiber Cross-Sectional Analysis

The 14-day muscle tissue samples were used for morphological analysis. Laminin staining was performed to determine the muscle fiber cross-sectional area (CSA), as previously described (18 (link), 22 (link)). Briefly, the TA muscle samples were obtained, fixed, and cut into 10-μm thick cryo-sections. Next, the sections were incubated with an anti-laminin antibody (Abcam plc., Cambridge, UK) at 4°C for 12 hours and afterward with a fluorescent secondary antibody (Invitrogen Alexa Fluor, Thermo-Fisher Scientific, Waltham, MA, USA) at room temperature for one hour. The sections were investigated and photographed under fluorescence microscopy (Zeiss Microscopy, Jena, Germany), and the muscle fiber CSA was determined using ImageJ software (National Institutes of Health, Bethesda, MD, USA).