Effects of O-GlcNAc inhibition on cell cycle and differentiation of mouse C2C12 cells
The efficient proliferation and differentiation of myoblasts are crucial for the production of livestock meat products.In this experiment,the specific inhibitor OSMI-1 was employed to inhibit O-linked β-N-acetylglucosamine(O-GlcNAc)glycosylation to observe the changes in the cell cycle and differentiation of the mouse myoblast cell line(C2C12).Mouse C2C12 cells were used as experimental subjects,with a blank group,a DMSO group,and an OSMI-1/DMSO group established.Western Blot was utilized to determine the inhibition of O-GlcNAc modification by OSMI-1;flow cytometry was used to detect changes in the cell cycle;immunofluorescence was used to detect the differentiation marker(myosin heavy chain,Myhc),and qRT-PCR was used to detect the expression levels of differentiation factors(myogenic differentiation,Myod)and Myhc genes,to analyze the effect of O-GlcNAc on C2C12 myotube differentiation.The results indicated that the O-GlcNAc modification level of C2C12 was significantly inhibited by the OSMI-1/DMSO group at24 h and 48 h,and the proportion of G2 cells in the cell cycle was significantly reduced(P<0.05).During the differentiation process,the inhibition of O-GlcNAc modification resulted in a decrease in the expression of the differentiation marker Myhc,as well as a decrease in the expression of Myod and Myhc.
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