Understanding how mitochondria are involved in myogenesis will provide a valuable insight into the underlying mechanisms that regulate the maintenance of cellular homeostasis. Recently, it has been reported that the transgenic mice with skeletal muscle-specific expression of PGC-1�� preserve mitochondrial function as well as neuromuscular junctions and muscle integrity during ageing [92], http://www.selleckchem.com/products/Belinostat.html and mitochondrial gene therapy may be effective in the treatment of muscle injury [58]. These efforts may facilitate to understand the molecular mechanisms of mitochondrial disorders.Figure 1Hypothetic model of mitochondrial activity in myogenic differentiation.AcknowledgmentThis research was supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) (Grant-in-Aid for Scientific Research (C), 22500658), Japan.

DENV-2 New Guinea C strain, recovered from the brain of newborn Swiss mice, was used in this study. The viral stock was prepared by inoculation of C6/36 cells contained in 75cm2 culture flasks with virus diluted in 1mL of L-15-2% FBS. After 1h, 14mL of L-15 supplemented with 10% FBS was added, and the cells were cultured for 7 days. Cells culture supernatant was then harvested and centrifuged at 2,000Xg for 5min to removed cell debris. The supernatant containing the virus was adjusted to 20% FBS, aliquoted, and stored at ?70��C. Virus stock and cell culture supernatants used in the present study were free of the lipopolysaccharide and mycoplasma.DENV-2 was used in the study because it showed the best of titration in preliminary experiments to assess the antiviral effect of chloroquine in Vero and C6/36 cells.

2.3. Dengue Virus TitrationVirus production was titrated by plaque assay using Vero cells. Vero cells were seeded in 12-well (6 �� 105 cells/well) plate in L-15 medium with 10% FBS for 48h at 37��C. Medium was removed, and decimal serial dilutions of virus stock or supernatant of cells treated with CLQ, prepared in L-15 medium with 2% FBS, were added (0.1mL/well) to the cells, which were then incubated for 2h at 37��C. Subsequently, L-15 medium containing 5% FBS and 3% carboxymethyl-cellulose (1mL/well) (overlay) was added, and the plate was incubated at 37��C for Entinostat 7 days. Overlay was removed on day seven, and cells were fixed with a solution of 10% formaldehyde in PBS. After 2 hours at room temperature, the formaldehyde solution was removed, and cells were washed twice with PBS and stained (15min) with a 1% crystal violet solution in 20% ethanol. The plaques of cell lysis were counted, and the virus concentration was expressed as plaque forming unites (PFU) per milliliter.