The LIM-Only Protein FHL2 is Involved in Autophagy to Regulate the Development of Skeletal Muscle

BioRxiv : the Preprint Server for Biology
Zihao LiuHuadong Yin

Abstract

Four and a half LIM domain protein 2 (FHL2) is a LIM domain protein expressed in muscle tissue whose deletion is causative of myopathies. Although FHL2 has a confirmed important role in muscle development, its autophagy-related function in muscle differentiation has not been fully determined. To explore the role of FHL2 in autophagy-related muscle regulation, FHL2-silenced and -overexpressing C2C12 mouse cells were examined. Immunofluorescence and co-immunoprecipitation assay findings showed that FHL2 silencing reduced LC3-II; protein expression and the amount of LC3 that co-immunoprecipitated with FHL2, indicating that FHL2 interacts with LC3-II; in the formation of autophagosomes. Moreover, the expression of muscle development marker genes such as MyoD1 and MyoG was lower in FHL2-silenced C2C12 cells but not in FHL2-overexpressing C2C12 cells. Electron microscopy analysis revealed large empty autophagosomes in FHL2-silenced myoblasts, while flow cytometry suggested that FHL2 silencing made cells more vulnerable to staurosporine-induced cell death. In conclusion, we propose that FHL2 interacts with LC3-II; in autophagosome formation to regulate the development of muscle cells.

Related Concepts

Autophagy
Cell Death
Flow Cytometry
Gene Deletion
Electron Microscopy
Muscle
Myopathy
Staurosporine
Immunofluorescence Assay
Skeletal Muscle Structure

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