Exosomes derived from inflammatory myoblasts promote M1 polarization and break the balance of myoblast proliferation/differentiation

Figure 2 Purification, isolation, and characterization of C2C12-Exos. A: Flowchart of C2C12-Exos purification based on differential ultra-centrifugation. Lipopolysaccharide, 1000 ng/mL for 1 d; B: The morphology of C2C12-Exos was observed by transmission electron microscopy (left) and scanning electron microscopy (right). The red arrows indicate representative exosomes; C: Representative flow cytometry plots showing the phenotypes of exosome markers, including CD9, CD47, CD63, and CD81; D: The particle size distribution of C2C12-Exos was analyzed by the qNano platform; E: Western blotting showed the presence of exosomal markers, including CD63, HSP60, Alix, and CD9. The four lanes represent different exosomal proteins and deproteinized supernatants extracted from the two independent conditioned media. LPS: Lipopolysaccharide.