Abstract

Simple SummaryThe purpose of this study was to initially determine the role of methylation of the promoter region of Forkhead box O 1 (FOXO1) in regulating its transcriptional level and to further investigate the effect ofFOXO1on the proliferation and differentiation of bovine myogenic cells. In this study, we used bisulfite sequencing polymerase chain reaction, real-time quantitative PCR, western blot, cell counting kit-8 (CCK-8), and flow cytometry and found that the mRNA expression of theFOXO1was low when the methylation ofFOXO1promoter region was high, and silencing the expression of theFOXO1gene could promote the proliferation and differentiation of myoblasts.AbstractThis study aimed to explore the regulatory role ofFOXO1promoter methylation on its transcriptional level and unravel the effect ofFOXO1on the proliferation and differentiation of bovine myoblasts. Bisulfite sequencing polymerase chain reaction (BSP) and real-time quantitative PCR were performed to determine the methylation status and transcript levels of theFOXO1promoter region at different growth stages. BSP results showed that the methylation level in the calf bovine (CB) group was significantly higher than that in the adult bovine (AB) group (p< 0.05). On the other hand, qRT-PCR results indicated that the mRNA expression level in the AB group was significantly higher than that in the CB group (p< 0.05), suggesting a significant decrease in gene expression at high levels of DNA methylation. CCK-8 and flow cytometry were applied to determine the effect of silencing theFOXO1gene on the proliferation of bovine myoblasts. Furthermore, qRT-PCR and Western blot were conducted to analyze the expression of genes associated with the proliferation and differentiation of bovine myoblasts. Results from CCK-8 revealed that the short hairpinFOXO1(shFOXO1) group significantly promoted the proliferation of myoblas