2.9. Surface Sensing of Translation (SUnSET) Assay

Protein synthesis was measured using the SUnSET technique, as previously described [20]. Briefly, C2C12 myoblasts were differentiated for 5 days. After 5 days, the cells were treated with or without 10 μM DEX and 20 μM SNA for 12 h, followed by 1 μM puromycin (P8833, Sigma-Aldrich, MO, USA) and incubated for 30 min. Puromycin-labelled proteins were assessed using western blotting with an anti-puromycin antibody (MABE343, Sigma-Aldrich, MO, USA) using an equal amount of total protein per sample.