Objective: To investigate the role of nesprin-1 in mouse embryonic stem cells differentiation into cardiomyocyte.
Methods: Hanging drop-suspension-adherence method was applied for the differentiation of mouse embryonic stem cells into cardiomyocytes under the inducing of salvia miltorrhiza and 5-azacytidine. Changes in nesprin-1 gene expression were detected by using Western blotting and immunofluorescent assay. RNA interference was used to reduce nesprin-1 protein levels to further investigate the importance of nesprin-1 in mouse embryonic stem cells differentiation, group I (target sequence AAAGCCAAGCACGCAACTA), group II (target sequence GGGAACCAACAGTGAGATT), group III (target sequence ACCAGGACATTGCGTACTA), and group IV (control group).
Results: The nesprin-1 isoform profile was altered in mouse embryonic stem cells differentiation. The rates of differentiation of the four groups were (17.78 +/- 1.92)%, (36.67 +/- 3.34)%, (44.42 +/- 5.08)%, (77.78 +/- 1.92)%; The rate of differentiation of group IV was higher than RNAi groups and the difference was significant (P < 0.05). In addition, compared with the control group, myosin in RNAi groups were dramatically reduced.
Conclusion: Nesprin-1 played important roles in mouse embryonic stem cells differentiation into cardiomyocyte. Nesprin-1 isoforms might perform different functions in the process of mouse embryonic stem cells differentiation.
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