Journal
INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES
Volume 15, Issue 4, Pages 838-846Publisher
IVYSPRING INT PUBL
DOI: 10.7150/ijbs.31371
Keywords
FHL2; LIM-domain; autophagy; cell development; C2C12 cells
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Funding
- China Agriculture Research System [CARS-40]
- Thirteenth Five Year Plan for Breeding Program in Sichuan [2016NYZ0050]
- Edanz Group China
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Scope: 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. Methods: C2C12 cells were treated with FHL2-konwdown or FHL2-overexpression. The morphology of C2C12 cells was observed by transmission electron microscopy. The mRNA and protein abundances of muscle related genes and autophagy related genes were measured by RT-PCR and western blot. Immunofluorescence and co-immunoprecipitation assay were used to verify the interaction between FHL2 and LC3 protein. Results: 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. Conclusion: These results suggest that FHL2 interacts with LC3- II in autophagosome formation to regulate the development of muscle cells.
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