Fig 1.
Differentiation of C2C12 and primary myoblast.
(A) The representative phase contrast images of C2C12 cells and primary myoblasts under induced differentiation at different time intervals (Day 0, 4 and 6). White arrows indicating the myotube formation. (B) Immunoblot analyses showing the expression profile of myogenesis and mitochondrial biogenesis markers of C2C12 and primary myoblasts under induced differentiation (p-STAT3, phosphorylated STAT3; Ac, acetylated STAT3 and t-STAT3, total STAT3).
Fig 2.
The cytotoxicity of GMI on C2C12 myoblasts.
(A) C2C12 cells were treated with different dosage of GMI (0, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20 and 30 μg/ml) for 24 hours and the morphological changes were visualized. (B) The cell viability of GMI pre-treated C2C12 was determined by using CCK-8 assay. Statistical analyses were performed by One-way ANOVA. ***, p < 0.001 and **** p < 0.0001.
Fig 3.
Promoted C2C12 myogenic differentiation with GMI pretreatment.
(A) The representative phase contrast images of induced differentiated C2C12 with pretreatment of GMI (0, 0.01, 0.05, 0.1, 0.5, 1, 5,10 and μg/ml) on day 6. Immunoblot analyses showing the expression profile of myogenesis, mitochondrial biogenesis and pro-inflammatory markers of GMI treated C2C12 (B) crude cells lysate. (C) IL-6 secretion from C2C12 cells with GMI pre-treatment afterward 24, 48 and 72 hours was determined by ELISA assays. Data are means ± SEM. *P<0.05, **, p < 0.01 ***, p < 0.001 and **** p < 0.0001.
Fig 4.
Schematic depicts that GMI pretreatment promotes C2C12 myogenic differentiation via activation of Tid1 and Ac-STAT3.