EPA ameliorates the effects of TNF-α on differentiating myotube appearance and MyHC expression. C2C12 cells were induced to differentiate in DM in the presence or absence of TNF-α (20 ng/ml). EPA (50 μM) was added together with TNF-α as a co-treatment (EPA). Alternatively, for some experiments EPA was administered alone for a 2 hour pre-treatment (pEPA) after which it was withdrawn and replaced by TNF-α alone in DM. Incubations were continued for 5 days in DM with replenishment of EPA and TNF-α at media changes. Representative images show immunofluorescence detection of Alexa-Fluor546 conjugated anti-MF20 antibody against MyHC (pink) and DAPI counterstained nuclei (blue) for various treatments with EPA and TNF-α. Control treatments are shown in the left panel images and TNF-α treatments in the right panel images. Calibration bars are 100 μm.