Priority Research Paper Volume 12, Issue 12 pp 11200—11223

Endothelin-1 induces cellular senescence and fibrosis in cultured myoblasts. A potential mechanism of aging-related sarcopenia

Figure 2. Endothelin-1 increases FN expression in mouse myoblasts (C2C12) through ETA receptor. Cells were incubated with 1 nM ET-1 at different times. Then, FN mRNA expression was assessed by RT-qPCR (panel A) and FN protein content by Western blot (panel B). To study the ET receptor implicated, cells were incubated with 100 nM BQ-123 (BQ123) or 100 nM BQ-788 (BQ788) added 30 min before ET-1 (1 nM), and then incubated for 24h. Then, FN protein content (panel C) as well as intracellular FN (in red) and extracellular FN (in green) expression (panel D) were studied by Western and immunofluorescence, respectively. In the experiments of the analysis of protein content, a representative Western blot is shown at the top with the densitometric analysis below (panel B,C). Values are the mean±SEM of 6 independent experiments, *p<0.05 vs. control cells (C), and **p<0.05 vs ET alone.