METTL14-Mediated m6a modification serves as an ERK downstream effector regulating myogenesis in alpas cashmere goat muscle stem cells

N⁶-methyladenosine (m⁶A) is the most abundant internal modification in eukaryotic mRNAs and plays critical roles in skeletal muscle development by regulating gene expression post-transcriptionally. However, the involvement of the m⁶A methyltransferase METTL14 in myogenesis of Alpas cashmere goats remains largely unexplored. Alpas cashmere goats are economically important livestock, and understanding the molecular mechanisms governing their muscle growth is of agricultural significance. The expression level of the m 6 A methyltransferase METTL14 in Alpas cashmere goat MuSCs was altered using overexpression and knockdown. The influence of METTL14 on the methylation levels, growth characteristics, and degree of the myogenic differentiation of MuSCs was measured using dot blot and Cell Counting Kit-8 assays, propidium iodide (PI) staining, Annexin V-FITC/PI staining, real-time quantitative polymerase chain reaction (RT-qPCR), and western blotting. The extracellular signal-regulated kinase (ERK) inhibitor PD0325901 was supplemented to the culture medium of MuSCs to determine whether METTL14 regulates MuSC growth and differentiation through the ERK signaling pathway. Myogenic differentiation was suppressed in METTL14-overexpressing MuSCs and promoted in METTL14 knockdown MuSCs. Both METTL14 overexpression and knockdown were accompanied by corresponding changes in global m⁶A methylation levels and ERK phosphorylation status. The ERK pathway inhibitor significantly promoted the protein expression of the myogenic differentiation marker genes MYOG and MYOD1 in MuSCs. METTL14 overexpression attenuated the degree to which the ERK pathway inhibitor promoted myogenic differentiation. The ERK protein regulates the myogenic differentiation of Alpas cashmere goat MuSCs through the m⁶A methyltransferase METTL14. These findings reveal a novel post-transcriptional regulatory mechanism in caprine skeletal muscle development and may have implications for improving meat production traits in livestock breeding.