IL-6 Impairs Myogenic Differentiation by Downmodulation of p90RSK/eEF2 and mTOR/p70S6K Axes, without Affecting AKT Activity
Figure 2
C-26 CM and exogenous IL-6 impair the myogenic differentiation of C2C12 cells. C2C12 myoblasts were cultured in GM until subconfluence and then shifted to differentiation medium (DM), or DM containing 10% C-26 CM, or DM containing 25 ng/mL of recombinant IL-6. (a) After 48 h in the differentiation media, cells were stained using an anti-sarcomeric myosin heavy chain (MF-20) antibody (red) and the Hoechst staining of the DNA (blue) and was imaged by conventional fluorescent microscopy. (b) After 48 h in the differentiation media, cells were harvested and total protein was extracted; cell lysates (70 μg of total protein) were subjected to Western blot analysis with the indicated specific antibodies. The experiment shown is representative of three independent determinations. (c) Densitometric analysis of the relative protein levels (obtained from Western blotting analysis) of sarcomeric myosin heavy chain, represented as arbitrary units (A.U.) and equalized for α-tubulin expression. (Student’s -test).
