Hesperetin but not Ellagic Acid Increases Myosin Heavy Chain Expression and Cell Fusion of Oxidative Stress Treated C2C12 Myoblasts

In this in vitro cell experiment, C2C12 myoblasts were proliferated for 24h and differentiated for 120h while exposed to different concentrations of hesperetin (5, 20, 50μM), ellagic acid (0.05, 0.1μM) or a combination (20μM hesperetin, 0.05μM, 20 ellagic acid) in the absence and presence of oxidative stress-inducing compound menadione (9μM) during 24h of proliferation and during the first 5h of differentiation.

The number of proliferating cells was assessed using fluorescent labelling of incorporated 5-ethynyl-2’-deoxyuridine (EdU). Myosin heavy chain (MyHC) expression was assessed by fluorescence microscopy and muscle cell fusion index was calculated as the number of nuclei inside MyHC-positive myotubes as a percentage of the total number of nuclei. Furthermore, protein expression of phosphorylated p38 and myomixer were assessed using Western blot.