Research Paper Volume 13, Issue 19 pp 22985—23003

Figure 5. VTG2 promotes the proliferation of C2C12 cells. (A) CCK-8 assay suggests that when VTG2 (5 ng/mL) is used to treat the cells for 36 h, they show significant proliferation. Each experiment was performed in quadruplicate wells; (B) Luciferase method is used in C2C12 cells. VTG2 can inhibit the activity of the MSTN promoter. Each experiment was performed in triplicate wells; (C) VTG2 (5 ng/mL) with the same concentration is used to treat C2C12 cells for different times (0 h, 12 h, 36 h, 48 h). Western blot assay suggests that VTG2 can promote the protein expression of MYOD, MYOG, and Desmin in C2C12 cells and inhibit the protein expression of muscle hydrolytic factors MSTN, Murf-1, and Atrogin-1. Each experiment was performed in triplicate wells; (D) During the differentiation of C2C12 cells, VTG2 (5 ng/mL) was added and treated for 36 hours. Immunofluorescence staining showed that VTG2 can promote myotube formation, (Bar = 100 μm); (E) VTG2 promotes the formation of lipid droplets; (F) VTG2 (5 ng/mL) is used to treat 3T3-L1 cells for 12 h, and the Western blot assay suggests that VTG2 can promote the protein expression of UCP1 and PGC1α in 3T3-L1 cells. Each experiment was performed in triplicate wells, (Bar = 200 μm); ^*P < 0.05, ^**P < 0.01.