Journal: Molecular Medicine Reports

Article Title: Succinate promotes skeletal muscle protein synthesis via Erk1/2 signaling pathway

doi: 10.3892/mmr.2017.7554

Figure Lengend Snippet: Succinate induced protein synthesis in C2C12 myotubes was mediated by Erk signaling pathway. C2C12 cells were cultured in a differentiation medium for 6 days. (A) [Ca 2+ ]i in C2C12 cells in response to succinate (20 mM). (B-D) C2C12 myotubes were exposed to succinate (0.5 and 2 mM) for 48 h. Western blot analysis of p-Akt and p-Erk. (E) The binding of Erk with Akt was tested by co-Immunoprecipitation. (F and G) Erk inhibitor U0126 (10 µM) was co-treated with succinate (2 mM) for 48 h. Puromycin levels were detected. Data are presented as mean ± SEM. a,b Significant differences between groups (P<0.05). *P<0.05 compared with the control or succinate group. β-actin was used as a loading control.

Article Snippet: C2C12 cell were treated with 0.5 and 2 mM succinate for 48 h. Cells were washed twice with cold PBS and lysed using 200 µl radio immunoprecipitation assay (RIPA) lysis buffer that contained protein phosphatase inhibitor complex (Biosino Bio-Technology and Science Inc., Beijing, China) and 1 mM PMSF.

Techniques: Cell Culture, Western Blot, Binding Assay, Immunoprecipitation