Key result
Knockdown of Ifitm1-3 by specific siRNAs significantly inhibited myogenesis of C2C12 myoblasts, decreasing the fusion index to 14.82% (P < 0.01) and downregulating differentiation factors.
p-value: p=<0.01
IFITM1-3 are upregulated during myogenic differentiation and their knockdown inhibits myogenesis, suggesting a role in muscle development and potential therapeutic targets for myopathic diseases.
IFITM upregulation during myoblast differentiation is hypothesis-generating; extends myogenesis research but requires functional validation.
Interferon-induced transmembrane proteins (IFITMs 1, 2, and 3) play a critical role in preventing pathogen infection in vertebrates. They are also involved in the occurrence and prognosis of cancer. Myogenesis is a complex process regulated by several factors. This study disclosed that Ifitm1-3 were upregulated in the process of myogenic differentiation of C2C12 myoblasts on days 3, 5, and 7. This positively correlated with the expression of differentiation factors MyoD, myogenin, Mrf5, and desmin. Furthermore, knockdown of Ifitm1-3 by their individual siRNAs inhibited myogenesis of C2C12 myoblasts, with relative downregulation of MyoD, myogenin, Mrf5, and desmin. Subsequently, myotube formation and fusion percentage decreased. Co-immunoprecipitation combined with LC-MS/MS analysis uncovered the interaction proteins of IFITM1 and IFITM3 in C2C12 myoblasts. A total of 84 overlapped interaction proteins of IFITM1 and IFITM3 were identified, and one of the clusters was engaged in cytoskeletal and sarcomere proteins, including desmin, myosin, actin, vimentin, nestin, ankycorbin, and nucleolin. Hence, we hypothesize that these interacting proteins may function as scaffolds for IFITM1-3, possibly through the interaction protein desmin to initiate further interaction with other proteins to participate in myogenesis; however, the molecular mechanisms remain unclear. Our study may contribute to the development of novel therapeutics for myopathic diseases.
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Zhang et al. (2023) studied Myogenic differentiation. siRNA targeting Ifitm1-3 vs. Mock / negative control siRNA was evaluated on Myotube formation and fusion percentage (fusion index) (p=<0.01). Knockdown of Ifitm1-3 by specific siRNAs significantly inhibited myogenesis of C2C12 myoblasts, decreasing the fusion index to 14.82% (P < 0.01) and downregulating differentiation factors.
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