Goat anti pax7

The human myogenic cells were isolated, as previously described35 (link). The FACS analysis was performed using goat anti-Pax7 (LifeSpan BioSciences, Seattle, WA, USA), rabbit anti-PTH1R (Sigma-Aldrich), and PE-Cy7-labeled anti-human CD34 (BioLegend, San Diego, CA, USA) primary antibodies. Alexa-Fluor-488-labeled donkey anti-goat IgG (Life Technologies) and PE-labeled donkey anti-rabbit IgG (eBioscience, San Diego, CA, USA) antibodies were used as secondary antibodies for Pax7 and PTH1R detection, respectively. Flow cytometry and cell sorting were performed using a BD FACSVantage SE flow cytometer (BD Bioscience, San Jose, CA, USA) commission to ReproCell (Kanagawa, Japan).

Human muscle biopsy specimens were obtained as described above. Samples of the gastrocnemius were collected from C57BL/10 and mdx mice following euthanization, and the tissues were cryosectioned. Hematoxylin and eosin (HE) staining was performed for the myofiber analysis. Rabbit anti-PTH1R (1:400; Sigma-Aldrich), goat anti-Pax7 (1:300; LifeSpan Bioscience), goat anti-CD34 (1:100; Santa Cruz Biotechnologies, Dallas, TX, USA), and goat anti-MyoD (1:100; Santa Cruz Biotechnologies) monoclonal antibodies were used as primary antibodies for immunohistochemical staining. Primary antibody reactivity was detected using an Alexa-Fluor-555-labeled ant-rabbit IgG (Life Technologies) or an Alexa-Fluor-488-labeled anti-goat IgG secondary antibody. The slides were examined using fluorescence microscopy.