Research Paper Volume 13, Issue 19 pp 22985—23003

Figure 2. FEYE promotes the proliferation and differentiation of skeletal muscle. (A) FEYE can effectively inhibit the mRNA expression of MSTN, Murf-1, and Atrogin-1 in skeletal muscle and significantly enhance mRNA expression of myocyte growth factor MYOD. Each experiment was performed in triplicate wells; (B) Western blot assay shows that FEYE can significantly inhibit the protein expression of MSTN in skeletal muscle and promote the expression of MYOD. Each experiment was performed in triplicate wells; (C) ELISA assay shows that FEYE can decrease the expression level of of MSTN in serum. Each experiment was performed in triplicate wells; (D) Western blot assay shows that FEYE can significantly enhance the phosphorylation of AKT. (E) CCK-8 assay is used to detect the proliferation rate of C2C12 cells; (F) FEYE (10 mg/mL) is added to produce action for 36 h during the proliferation of C2C12 cells. FEYE can promote cell proliferation; (G) FEYE is added in the proliferated and differentiated C2C12 cells at the concentration of 10 mg/mL. Immunofluorescence staining of eMyHC shows that FEYE can enhance the formation of myotubes; (Bar = 100 μm); ^*P < 0.05, ^**P < 0.01.