In vitro effects of 1α,25(OH)2D3-glycosides from Solbone A (Solanum glaucophyllum leaves extract; Herbonis AG) compared to synthetic 1α,25(OH)2D3 on myogenesis

1α,25(OH)2D3-glycosides fromSolanum glaucophyllumleaves extract induce myoblasts differentiation through p38 MAPK and AKT activation

“…C2C12 myoblasts cultured in these conditions resemble the activated satellite cells ( Yoshida et al, 1998 ). Myoblast differentiation was promoted as was previously reported ( Gili et al, 2016 ). Briefly, (with C2C12 cells at 70–80% of confluence) GM was replaced with DMEM without phenol red, supplemented with 2% HS (DM).…”

“…We used the murine skeletal muscle cell line C2C12 to study myogenesis ( Gili et al, 2016 ). Cells were cultured with DMEM supplemented with 10% heat-inactivated (30 min, 56°C) FBS, 1% nystatine, and 1% streptomycin, GM at 37°C under a humidified atmosphere of 5% CO 2 in air.…”

“…C2C12 cells were scraped in 75 µl of lysis buffer and lysates were collected by aspiration and centrifuged at 14,000 g for 15 min ( Gili et al, 2016 ). Proteins from supernatant were quantified using the Bradford procedure as before ( Gili et al, 2016 ; Bradford, 1976 ).…”

“…Proteins were separated on SDS-PAGE (10% gels) and electro transferred to PVDF membranes. Membranes were then blocked and subjected to immunoblotting ( Gili et al, 2016 ). Incubation with primary antibodies, (1:1000) in 0.1% TBST containing 2.5% dry milk, was performed overnight at 4°C.…”

“…We have previously demonstrated that SGE increases VDR, myogenin, and myosin heavy chain protein levels during the first 48 h of C2C12 muscle cell line differentiation ( Gili et al, 2016 ). Moreover, SGE improves the growth and differentiation of C2C12 cells at the onset of myogenesis, increasing cellular mass and myotube fusion ( Gili et al, 2016 ). In this work, we further studied the effects of SGE on the signaling pathways involved in C2C12 muscle cell differentiation.…”