EPA ameliorates baseline and TNF-α-induced cellular necrosis. Any potential cytotoxic effect of EPA on C2C12 myotube formation was investigated after 5 days following induction of differentiation by DM using a CellTiter Blue assay to detect all non-viable cells (A). The effect of TNF-α (20 ng/ml) on cellular necrosis and the ability of EPA to block this activity were also assessed by Trypan Blue dye exclusion to detect necrotic cells (B). Here, EPA was added together with TNF-α as a co-treatment (EPA+TNF) or, alternatively EPA was administered alone for a 2 hour pre-treatment after which it was withdrawn and replaced by TNF-α alone in DM (pEPA+TNF). Incubations were continued for 5 days in DM with replenishment of EPA and TNF-α at media changes. Data are expressed as means ± standard error of mean (SEM) from 3 independent experiments (*p < 0.05 v. control; #NS v. control; **p < 0.05 v. TNF-α).